DK2831111T3 - Ang2-binding molecules - Google Patents

Description

DESCRIPTION

FIELD OF THE INVENTION

[0001] The invention relates to the field of human therapy, in particular cancer therapy and agents and compositions useful in such therapy.

BACKGROUND OF THE INVENTION

[0002] As described in e.g. US 2008/0014196, WO 2008101985 and US 2011/0027286, angiogenesis is the biological process whereby new blood vessels are formed and being implicated in the pathogenesis of a number of disorders, including solid tumors and metastasis as well as eye diseases. When tumors reach a critical size of approximately 1 mm3 they become dependent on angiogenesis for maintaining blood supply with oxygen and nutritients to allow for further growth. Anti-angiogenesis therapies have become an important treatment option for several types of tumors.

[0003] One of the most important pro-angiogenic factors is Angiopoietin2 (Ang2), a ligand of the Tie2 receptor (Tie2), which controls vascular remodeling by enabling the functions of other angiogenic factors, such as VEGF. Ang2 is also referred to in the art as Tie2 ligand (US. Pat. No. 5,643,755, Yancopoulos et al. 2000, Nature 407: 242-248).

[0004] Ang2 is primarily expressed by endothelial cells, strongly induced by hypoxia and other angiogenic factors and has been demonstrated to regulate tumor vessel plasticity, allowing vessels to respond to VEGF and FGF2 (Augustin et al., Nat Rev Mol Cell Biol. 2009 Mar;10(3):165-77). Consistent with this role, the deletion or inhibition of Ang2 results in reduced angiogenesis (Falcon et al., Am J Pathol. 2009 Nov;175(5):2159-70). Elevated Ang2 serum concentrations have been reported for patients with colorectal cancer, NSCLC and melanoma (Goede et al., Br J Cancer. 2010 Oct 26;103(9):1407-14),(Park et al., Chest. 2007 Jul;132(1): 200-6),(Helfrich et al., Clin Cancer Res. 2009 Feb 15; 15(4): 1384-92). In CRC cancer Ang2 serum levels correlate with therapeutic response to anti-VEGF therapy.

[0005] The Ang-Tie system consists of 2 receptors (Tie 1 and Tie2) and 3 ligands (Ang1, Ang2 and Ang4) (Augustin et al., Nat Rev Mol Cell Biol. 2009 Mar;10(3):165-77). Tie2, Ang1 and Ang2 are the best studied members of this family, Tie1 is an orphan receptor and the role of Ang4 for vascular remodelling still needs to be defined. Ang2 and Ang1 mediate opposing functions upon Tie2 binding and activation. Ang2-mediated Tie2 activation results in endothelial cell activation, pericyte dissociation, vessel leakage and induction of vessel sprouting. In contrast to Ang2, Ang1 signaling maintains vessel integrity by recruitment of pericytes, thereby maintaining endothelial cell quiescence.

[0006] Angiopoietin 2 (Ang2) is a secreted, 66 kDa ligand for the Tie2 receptor tyrosine kinase (Augustin et al., Nat Rev Mol Cell Biol. 2009 Mar; 10(3): 165-77). Ang2 consists of an N-terminal coiled-coil domain and a C-terminal fibrinogen-like domain, the latter is required for Tie2 interaction. Ang2 is primarily expressed by endothelial cells and strongly induced by hypoxia and other angiogenic factors, including VEGF. Tie2 is found on endothelial cells, haematopoietic stem cells and tumor cells. Ang2-Tie2 has been demonstrated to regulate tumor vessel plasticity, allowing vessels to respond to VEGF and FGF2.

[0007] In vitro Ang2 has been shown to act as a modest mitogen, chemoattractant and inducer of tube formation in human umbilical vein endothelial cells (HUVEC). Ang2 induces tyrosine phosphorylation of ectopically expressed Tie2 in fibroblasts and promotes downstream signaling events, such as phosphorylation of ERK-MAPK, AKT and FAK in HUVEC. An antagonistic role of Ang2 in Ang1-induced endothelial cell responses has been described.

[0008] Ang2 -deficiency has been shown to result in a profound lymphatic patterning defect in mice. Although the loss of Ang2 is dispensable for embryonic vascular development, Ang2 -deficient mice have persistent vascular defects in the retina and kidney. Together with the dynamic pattern of Ang2 expression at sites of angiogenesis (for example ovary), these findings indicate that Ang2 controls vascular remodeling by enabling the functions of other angiogenic factors, such as VEGF.

[0009] The Ang2-Tie2 system exerts crucial roles during the angiogenic switch and later stages of tumor angiogenesis. Ang2 expression is strongly up-regulated in the tumor-associated endothelium. Reduced growth of tumors has been observed when implanted into Ang2 -deficient mice, especially during early stages of tumor growth. Therapeutic blocking of Ang2 with Ang2 mAbs has shown broad efficacy in a variety of tumor xenograft models.

[0010] As summarized in the above mentioned US 2008/0014196, angiogenesis is implicated in the pathogenesis of a number of disorders, including solid tumors and metastasis.

[0011] In the case of tumor growth, angiogenesis appears to be crucial for the transition from hyperplasia to neoplasia, and for providing nourishment for the growth and metastasis of the tumor. Folkman et al., Nature 339 -58 (1989), which allows the tumor cells to acquire a growth advantage compared to the normal cells. Therefore, anti-angiogenesis therapies have become an important treatment option for several types of tumors. These therapies have focused on blocking the VEGF pathway (Ferrara et al., Nat Rev Drug Discov. 2004 May;3(5):391-400)).

[0012] Antibodies and other peptide inhibitors that bind to Ang2 and Ang1 are mentioned in e.g. U.S. Patent Nos 6,166,185; 7,521,053; 7,205,275; and US Patent Application Nos.: 2006/0018909 and 2006/0246071. Furthermore, US 2011/0027286 discloses specific monoclonal Ang2 antibodies which do not antagonize the related molecule Ang1.

[0013] However, the state-of-the art monoclonal antibodies (MAbs) and fusion proteins have several shortcomings in view of their therapeutic application: To prevent their degradation, they must be stored at near freezing temperatures. Also, since they are quickly digested in the gut, they are not suited for oral administration. Another major restriction of MAbs for cancer therapy is poor transport, which results in low concentrations and a lack of targeting of all cells in a tumor.

[0014] It has been an object of the present invention to provide novel improved Ang2-binding molecules, i.e. nanobodies that block binding of Ang2 to Tie2 but not the binding of Ang1 to Tie2.

[0015] More in particular, it has been object of the invention to provide novel Ang2- binding molecules, and, specifically, Ang2-binding molecules that bind to mammalian Ang2 but not to mammalian Ang1 and, especially, to human Ang2 but not to human Ang1, wherein such molecules or polypeptides are suitable for the therapeutic and diagnostic purposes as described herein. It has been a further object of the invention to provide immunoglobulin single variable domains that specifically bind to Ang2 but not to Ang1.

[0016] Such Ang2- binding molecules, or Ang2 antagonists, are useful as pharmacologically active agents in compositions in the prevention, treatment, alleviation and/or diagnosis of diseases or conditions associated with Ang2-mediated effects on angiogenesis.

[0017] Examples for such diseases are cancer or cancerous diseases such as breast cancer, renal cell carcinoma, ovarian cancer and pancreatic cancer, eye diseases such as age-related macular degeneration and diabetic retinopathy, and/or chronic kidney diseases such as diabetic nephropathy, postrenal failure, prerenal azotemia and intrinsic renal failure.

[0018] It has been a further object of the invention to provide methods for the prevention, treatment, alleviation and/or diagnosis of such diseases, disorders or conditions, involving the use and/or administration of such Ang2-binding molecules and compositions comprising them. In particular, it is has been an object of the invention to provide such pharmacologically active Ang2-binding molecules, compositions and/or methods that provide advantages compared to the agents, compositions and/or methods currently used and/or known in the art. These advantages include improved therapeutic and/or pharmacological properties and/or other advantageous properties, e.g. for manufacturing purposes, especially as compared to conventional antibodies as those described above, or fragments thereof.

BRIEF SUMMARY OF THE INVENTION

[0019] According to a first aspect, there is provided an Ang2-binding molecule, comprising an immunoglobulin single variable domain, wherein said immunoglobulin single variable domain comprises three complementarity determining regions CDR1, CDR2 and CDR3, wherein CDR1 has an amino acid sequence shown in SEQ ID NO: 168, CDR2 has an amino acid sequence shown in SEQ ID No: 171 and CDR3 has an amino acid sequence shown in SEQ ID NO: 175.

[0020] The invention further relates to an Ang2-binding molecule consisting of said immunoglobulin single variable domain.

[0021] Furthermore, the invention relates to an Ang2-binding molecule having a sequence according to SEQ ID No: 166.

[0022] According to another aspect, there is provided a nucleic acid encoding said Ang2-binding molecule as well as an expression vector comprising said nucleci acid.

[0023] The invention further relates to a host cell carrying one or more expression vectors comprising said nucleic acids.

[0024] The invention further relates to a method for producing or generating an Ang2-binding molecule according to the invention, comprising the steps of: 1. (a) transfecting a host cell with one or more said vectors comprising said nucleic acid molecule, 2. (b) culturing said host cell, and 3. (c) recovering and purifying said Ang2-binding molecule.

[0025] Further aspect of the invention relates to a pharmaceutical composition comprising, as the active ingredient, one or more said Ang2-binding molecules as defined above and at least a physiologically acceptable carrier [0026] The invention further relates to applications and uses of the Ang2-binding molecules, nucleic acids, host cells, products and compositions described above, as well as to methods for the prevention and/or treatment for diseases associated with Ang2-mediated effects on angiogenesis, preferably cancer, cancerous diseases and eye diseases using the Ang2-binding molecules as defined above.

[0027] These and other aspects, embodiments, advantages and applications of the invention will become clear from the further description hereinbelow.

DEFINITIONS

[0028] Unless indicated or defined otherwise, all terms used have their usual meaning in the art, which will be clear to the skilled person. Reference is for example made to the standard handbooks, such as Sambrook et al, "Molecular Cloning: A Laboratory Manual" (2nd Ed.), Vols. 1-3, Cold Spring Harbor Laboratory Press (1989); Lewin, "Genes IV", Oxford University Press, New York, (1990), and Roitt et al., "Immunology" (2nd Ed.), Gower Medical Publishing, London, New York (1989), as well as to the general background art cited herein. Furthermore, unless indicated otherwise, all methods, steps, techniques and manipulations that are not specifically described in detail can be performed and have been performed in a manner known per se, as will be clear to the skilled person. Reference is for example again made to the standard handbooks, to the general background art referred to above and to the further references cited therein.

[0029] The term "angiopoetin-2" or "Ang2", unless specified as being from non-human species (e.g. "mouse Ang2", etc) refers to human Ang2 or a biologically active fragment thereof (e.g. a fragment of the Ang2 protein which is capable of inducing angiogenesis in vitro or in vivo), i.e. to human Ang2 (variant 1) with accession no. NM 001147.2, to human Ang2 (variant 2) having accession no. NM_001118887.1 or to human Ang2 (variant 3) having accession no. NM_001118888.1, and/or their biologically active fragments thereof. The amino acid sequences of Ang2 non human species, such as mouse Ang2 and cyno Ang2 are avialable from protein sequence database under Accession No: NM_007426.3 (SEQ ID NO: (SEQ ID 188) and AB172643.1 (SEQ ID NO: 187), respectively.

[0030] The term "angiopoietin-1"or "Ang1", unless specified as being from non-human species (e.g. "mouse Ang2", etc) refers to human Ang1 or a biologically active fragment thereof (e.g. a fragment of the Ang1 protein which is capable of inducing angiogenesis in vitro or in vivo), i.e. to human Ang1 with accession no. NM_001146.3, or a biologically active fragment thereof.

[0031] Unless indicated otherwise, the terms "immunoglobulin"and "immunoglobulin sequence" - whether used herein to refer to a heavy chain antibody or to a conventional 4-chain antibody -are used as general terms to include both the full-size antibody, the individual chains thereof, as well as all parts, domains or fragments thereof (including but not limited to antigen-binding domains or fragments such as VHH domains or VH/VL domains, respectively). In addition, the term "sequence" as used herein (for example in terms like "immunoglobulin sequence", "antibody sequence", "(single) variable domain sequence", "VHH sequence" or "protein sequence"), should generally be understood to include both the relevant amino acid sequence as well as nucleic acid sequences or nucleotide sequences encoding the same, unless the context requires a more limited interpretation.

[0032] The term "domain" (of a polypeptide or protein) as used herein refers to a folded protein structure which has the ability to retain its tertiary structure independently of the rest of the protein. Generally, domains are responsible for discrete functional properties of proteins and in many cases may be added, removed or transferred to other proteins without loss of function of the remainder of the protein and/or of the domain.

[0033] The term "immunoglobulin domain" as used herein refers to a globular region of an antibody chain (such as e.g. a chain of a conventional 4-chain antibody or of a heavy chain antibody), or to a polypeptide that essentially consists of such a globular region. Immunoglobulin domains are characterized in that they retain the immunoglobulin fold characteristic of antibody molecules, which consists of a 2-layer sandwich of about 7 antiparallel beta-strands arranged in two beta-sheets, optionally stabilized by a conserved disulphide bond.

[0034] The term "immunoglobulin variable domain" as used herein means an immunoglobulin domain essentially consisting of four "framework regions" which are referred to in the art and hereinbelow as "framework region 1" or "FR1"; as "framework region 2" or"FR2"; as "framework region 3" or "FR3"; and as "framework region 4" or "FR4", respectively; which framework regions are interrupted by three "complementarity determining regions" or "CDRs", which are referred to in the art and hereinbelow as "complementarity determining region 1"or "CDR1"; as "complementarity determining region 2" or "CDR2"; and as "complementarity determining region 3" or "CDR3", respectively. Thus, the general structure or sequence of an immunoglobulin variable domain can be indicated as follows: FR1 - CDR1 - FR2 - CDR2 - FR3 -CDR3 - FR4. It is the immunoglobulin variable domain(s) that confer specificity to an antibody for the antigen by carrying the antigen-binding site.

[0035] The term "immunoglobulin single variable domain" as used herein means an immunoglobulin variable domain which is capable of specifically binding to an epitope of the antigen without pairing with an additional variable immunoglobulin domain. One example of immunoglobulin single variable domains in the meaning of the present invention are "domain antibodies", such as the immunoglobulin single variable domains VH and VL (VH domains and VL domains). Another example of immunoglobulin single variable domains are "VHH domains" (or simply "VHHs") from camelids, as defined hereinafter.

[0036] In view of the above definition, the antigen-binding domain of a conventional 4-chain antibody (such as an IgG, IgM, IgA, IgD or IgE molecule; known in the art) or of a Fab fragment, a F(ab')2 fragment, an Fv fragment such as a disulphide linked Fv or a scFv fragment, or a diabody (all known in the art) derived from such conventional 4-chain antibody, would normally not be regarded as an immunoglobulin single variable domain, as, in these cases, binding to the respective epitope of an antigen would normally not occur by one (single) immunoglobulin domain but by a pair of (associating) immunoglobulin domains such as light and heavy chain variable domains, i.e. by a VH-VL pair of immunoglobulin domains, which jointly bind to an epitope of the respective antigen.

[0037] "VHH domains", also known as VHHs, VhH domains, VHH antibody fragments, and VHH antibodies, have originally been described as the antigen binding immunoglobulin (variable) domain of "heavy chain antibodies" (i.e. of "antibodies devoid of light chains"; Hamers-Casterman C, Atarhouch T, Muyldermans S, Robinson G, Hamers C, Songa EB, Bendahman N, Hamers R.: "Naturally occurring antibodies devoid of light chains"; Nature 363, 446-448 (1993)). The term "VHH domain" has been chosen in order to distinguish these variable domains from the heavy chain variable domains that are present in conventional 4-chain antibodies (which are referred to herein as "Vh domains" or "VH domains") and from the light chain variable domains that are present in conventional 4-chain antibodies (which are referred to herein as "V|_ domains" or "VL domains"). VHH domains can specifically bind to an epitope without an additional antigen binding domain (as opposed to VH or VL domains in a conventional 4-chain antibody, in which case the epitope is recognized by a VL domain together with a VH domain). VHH domains are small, robust and efficient antigen recognition units formed by a single immunoglobulin domain.

[0038] In the context of the present invention, the terms VHH domain, VHH, VhH domain, VHH antibody fragment, VHH antibody, as well as "Nanobody®" and "Nanobody® domain" ("Nanobody" being a trademark of the company Ablynx N.V.; Ghent; Belgium) are used interchangeably and are representatives of immunoglobulin single variable domains (having the structure FR1-CDR1-FR2-CDR2-FR3-CDR3-FR4 and specifically binding to an epitope without requiring the presence of a second immunoglobulin variable domain), and which are distinguished from VH domains by the so-called "hallmark residues", as defined in e.g. W02009/109635, Fig. 1.

[0039] The amino acid residues of a immunoglobulin single variable domain, e.g. a VHH, are numbered according to the general numbering for Vh domains given by Kabat et al. ("Sequence of proteins of immunological interest", US Public Health Services, NIH Bethesda, MD, Publication No. 91), as applied to VHH domains from Camelids, as shown e.g. in Figure 2 of Riechmann and Muyldermans, J. Immunol. Methods 231,25-38 (1999).

[0040] According to this numbering, • FR1 comprises the amino acid residues at positions 1-30, • CDR1 comprises the amino acid residues at positions 31-35, • FR2 comprises the amino acids at positions 36-49, • CDR2 comprises the amino acid residues at positions 50-65, • FR3 comprises the amino acid residues at positions 66-94, • CDR3 comprises the amino acid residues at positions 95-102, and • FR4 comprises the amino acid residues at positions 103-113.

[0041] However, it should be noted that - as is well known in the art for Vh domains and for VHH domains - the total number of amino acid residues in each of the CDRs may vary and may not correspond to the total number of amino acid residues indicated by the Kabat numbering (that is, one or more positions according to the Kabat numbering may not be occupied in the actual sequence, or the actual sequence may contain more amino acid residues than the number allowed for by the Kabat numbering). This means that, generally, the numbering according to Kabat may or may not correspond to the actual numbering of the amino acid residues in the actual sequence.

[0042] Alternative methods for numbering the amino acid residues of Vh domains, which methods can also be applied in an analogous manner to VHH domains, are known in the art. However, in the present description, claims and figures, the numbering according to Kabat and applied to VHH domains as described above will be followed, unless indicated otherwise.

[0043] The total number of amino acid residues in a VHH domain will usually be in the range of from 110 to 120, often between 112 and 115. It should however be noted that smaller and longer sequences may also be suitable for the purposes described herein.

[0044] Immunoglobulin single variable domains, e.g. VHHs and domain antibodies, according to the preferred embodiments of the invention as defined above, have a number of unique structural characteristics and functional properties which makes them highly advantageous for use in therapy as functional antigen-binding molecules. In particular, and without being limited thereto, VHH domains (which have been "designed" by nature to functionally bind to an antigen without pairing with a light chain variable domain) can function as single, relatively small, functional antigen-binding structural units.

[0045] Due to their unique properties, immunoglobulin single variable domains, as defined herein, like VHHs or VHs (or VLs) - either alone or as part of a larger polypeptide, e.g. a biparatopic molecule - offer a number of significant advantages: • only a single domain is required to bind an antigen with high affinity and with high selectivity, so that there is no need to have two separate domains present, nor to assure that these two domains are present in the right spacial conformation and configuration (i.e. through the use of especially designed linkers, as with scFv's); • immunoglobulin single variable domains can be expressed from a single nucleic acid molecule and do not require any post-translational modification (like glycosylation); • immunoglobulin single variable domains can easily be engineered into multivalent and multispecific formats (as further discussed herein); • immunoglobulin single variable domains have high specificity and affinity for their target, low inherent toxicity and can be administered via alternative routes than infusion or injection; • immunoglobulin single variable domains are highly stable to heat, pH, proteases and other denaturing agents or conditions and, thus, may be prepared, stored or transported without the use of refrigeration equipments; • immunoglobulin single variable domains are easy and relatively inexpensive to prepare, both on small scale and on a manufacturing scale. For example, immunoglobulin single variable domains can be produced using microbial fermentation (e.g. as further described below) and do not require the use of mammalian expression systems, as with for example conventional antibodies; • immunoglobulin single variable domains are relatively small (approximately 15 kDa, or 10 times smaller than a conventional IgG) compared to conventional 4-chain antibodies and antigen-binding fragments thereof, and therefore show high(er) penetration into tissues (including but not limited to solid tumors and other dense tissues) and can be administered in higher doses than such conventional 4-chain antibodies and antigenbinding fragments thereof; • VHHs have specific so-called "cavity-binding properties" (inter alia due to their extended CDR3 loop, compared to VH domains from 4-chain antibodies) and can therefore also access targets and epitopes not accessible to conventional 4-chain antibodies and antigen-binding fragments thereof; • VHHs have the particular advantage that they are highly soluble and very stable and do not have a tendency to aggregate (as with the mouse-derived antigen-binding domains described by Ward etal., Nature 341: 544-546 (1989)).

[0046] The immunoglobulin single variable domains of the invention are not limited with respect to a specific biological source from which they have been obtained or to a specific method of preparation. For example, obtaining VHHs may include the following steps: 1. (1) isolating the VHH domain of a naturally occurring heavy chain antibody; or screening a library comprising heavy chain antibodies or VHHs and isolating VHHs therefrom; 2. (2) expressing a nucleic acid molecule encoding a VHH with the naturally occurring sequence; 3. (3) "humanizing" (as described herein) a VHH, optionally after affinity maturation, with a naturally occurring sequence or expressing a nucleic acid encoding such humanized VHH; 4. (4) "camelizing" (as described below) a immunoglobulin single variable heavy domain from a naturally occurring antibody from an animal species, in particular a species of mammal, such as from a human being, or expressing a nucleic acid molecule encoding such camelized domain; 5. (5) "camelizing" a VH, or expressing a nucleic acid molecule encoding such a camelized VH; 6. (6) using techniques for preparing synthetically or semi-synthetically proteins, polypeptides or other amino acid sequences; 7. (7) preparing a nucleic acid molecule encoding a VHH domain using techniques for nucleic acid synthesis, followed by expression of the nucleic acid thus obtained; 8. (8) subjecting heavy chain antibodies or VHHs to affinity maturation, to mutagenesis (e.g. random mutagenesis or site-directed mutagenesis) and/or any other technique(s) in order to increase the affinity and/or specificity of the VHH; and/or 9. (9) combinations or selections of the foregoing steps.

[0047] Suitable methods and techniques for performing the above-described steps are known in the art and will be clear to the skilled person. By way of example, methods of obtaining VHH domains binding to a specific antigen or epitope have been described in W02006/040153 and W02006/122786.

[0048] According to specific embodiments, the immunoglobulin single variable domains of the invention or present in the polypeptides of the invention are VHH domains with an amino acid sequence that essentially corresponds to the amino acid sequence of a naturally occurring VHH domain, but that has been "humanized" or "sequence-optimized" (optionally after affinity-maturation), i.e. by replacing one or more amino acid residues in the amino acid sequence of said naturally occurring VHH sequence by one or more of the amino acid residues that occur at the corresponding position(s) in a variable heavy domain of a conventional 4-chain antibody from a human being. This can be performed using methods known in the art, which can by routinely used by the skilled person.

[0049] A humanized VHH domain may contain one or more fully human framework region sequences, and, in an even more specific embodiment, may contain human framework region sequences derived from the human germline Vh3 sequences DP-29, DP-47, DP-51, or parts thereof, or be highly homologous thereto, optionally combined with JH sequences, such as JH5. Thus, a humanization protocol may comprise the replacement of any of the VHH residues with the corresponding framework 1, 2 and 3 (FRI, FR2 and FR3) residues of germline VH genes such as DP 47, DP 29 and DP 51) either alone or in combination. Suitable framework regions (FR) of the immunoglobulin single variable domains of the invention can be selected from those as set out e.g. in WO 2006/004678 and specifically, include the so-called "KERE" and "GLEW" classes. Examples are immunoglobulin single variable domains having the amino acid sequence G-L-E-W at about positions 44 to 47, and their respective humanized counterparts. A humanized VHH domain may contain one or more fully human framework region sequences.

[0050] By way of example, a humanizing substitution for VHHs belonging to the 103 P,R,S-group and/or the GLEW-group (as defined below) is 108Q to 108L. Methods for humanizing immunoglobulin single variable domains are known in the art.

[0051] Binding immunoglobulin single variable domains with improved properties in view of therapeutic application, e.g. enhanced affinity or decreased immunogenicity, may be obtained from individual binding molecules by techniques known in the art, such as affinity maturation (for example, starting from synthetic, random or naturally occurring immunoglobulin sequences), CDR grafting, humanizing, combining fragments derived from different immunoglobulin sequences, PCR assembly using overlapping primers, and similar techniques for engineering immunoglobulin sequences well known to the skilled person; or any suitable combination of any of the foregoing, also termed "sequence optimization", as described herein. Reference is, for example, made to standard handbooks, as well as to the further description and Examples.

[0052] If appropriate, a binding molecule with increased affinity may be obtained by affinity-maturation of another binding molecule, the latter representing, with respect to the affinity-matured molecule, the "parent" binding molecule.

[0053] Methods of obtaining VHHs that bind to a specific antigen or epitope have been described earlier, e.g. in W02006/040153 and W02006/122786. As also described therein in detail, VHH domains derived from camelids can be "humanized" (also termed "sequence-optimized" herein, "sequence-optimizing" may, in addition to humanization, encompass an additional modification of the sequence by one or more mutations that furnish the VHH with improved properties, such as the removal of potential post translational modification sites) by replacing one or more amino acid residues in the amino acid sequence of the original VHH sequence by one or more of the amino acid residues that occur at the corresponding position(s) in a VH domain from a conventional 4-chain antibody from a human being. A humanized VHH domain can contain one or more fully human framework region sequences, and, in an even more specific embodiment, can contain human framework region sequences derived from DP-29, DP-47, DP-51, or parts thereof, optionally combined with JH sequences, such as JH5.

[0054] "Domain antibodies", also known as "Dab"s and "dAbs" (the terms "Domain Antibodies" and "dAbs" being used as trademarks by the GlaxoSmithKline group of companies) have been described in e.g. Ward, E.S., et al.: "Binding activities of a repertoire of single immunoglobulin variable domains secreted from Escherichia coli"; Nature 341: 544-546 (1989); Holt, L.J. et al.: "Domain antibodies: proteins for therapy"; TRENDS in Biotechnology 21(11): 484-490 (2003); and W02003/002609.

[0055] Domain antibodies essentially correspond to the VH or VL domains of antibodies from non-camelid mammals, in particular human 4-chain antibodies. In order to bind an epitope as a single antigen binding domain, i.e. without being paired with a VL or VH domain, respectively, specific selection for such antigen binding properties is required, e.g. by using libraries of human single VH or VL domain sequences.

[0056] Domain antibodies have, like VHHs, a molecular weight of approximately 13 to approximately 16 kDa and, if derived from fully human sequences, do not require humanization for e.g. therapeutical use in humans. As in the case of VHH domains, they are well expressed also in prokaryotic expression systems, providing a significant reduction in overall manufacturing cost.

[0057] Furthermore, it will also be clear to the skilled person that it is possible to "graft" one or more of the CDR's mentioned above onto other "scaffolds", including but not limited to human scaffolds or non-immunoglobulin scaffolds. Suitable scaffolds and techniques for such CDR grafting are known in the art.

[0058] The terms "epitope" and "antigenic determinant", which can be used interchangeably, refer to the part of a macromolecule, such as a polypeptide that is recognized by antigenbinding molecules, such as conventional antibodies or the polypeptides of the invention, and more particularly by the antigen-binding site of said molecules. Epitopes define the minimum binding site for an immunoglobulin, and thus represent the target of specificity of an immunoglobulin.

[0059] A polypeptide (such as an immunoglobulin, an antibody, an immunoglobulin single variable domain of the invention, or generally an antigen-binding molecule or a fragment thereof) that can "bind to" or "specifically bind to", that "has affinity for" and/or that "has specificity for" a certain epitope, antigen or protein (or for at least one part, fragment or epitope thereof) is said to be "against" or "directed against" said epitope, antigen or protein or is a "binding" molecule with respect to such epitope, antigen or protein. In this context, an Ang2-binding molecule may also be referred to as "Ang2-neutralizing".

[0060] Generally, the term "specificity" refers to the number of different types of antigens or epitopes to which a particular antigen-binding molecule or antigen-binding protein (such as an immunoglobulin single variable domain of the invention) molecule can bind. The specificity of an antigen-binding molecule can be determined based on its affinity and/or avidity. The affinity, represented by the equilibrium constant for the dissociation of an antigen with an antigenbinding protein (KD), is a measure for the binding strength between an epitope and an antigenbinding site on the antigen-binding protein: the lesser the value of the KD, the stronger the binding strength between an epitope and the antigen-binding molecule (alternatively, the affinity can also be expressed as the affinity constant (KA), which is 1/KD). As will be clear to the skilled person (for example on the basis of the further disclosure herein), affinity can be determined in a manner known per se, depending on the specific antigen of interest. Avidity is the measure of the strength of binding between an antigen-binding molecule (such as an immunoglobulin, an antibody, an immunoglobulin single variable domain or a polypeptide containing it and the pertinent antigen. Avidity is related to both the affinity between an epitope and its antigen binding site on the antigen-binding molecule and the number of pertinent binding sites present on the antigen-binding molecule.

[0061] The part of an antigen-binding molecule (such as an antibody or an immunoglobulin single variable domain of the invention) that recognizes the epitope is called a "paratope".

[0062] Unless indicated otherwise, the term "Ang2-binding molecule" includes anti- Ang2 antibodies, anti- Ang2 antibody fragments, "anti- Ang2 antibody-like molecules" and conjugates with any of these. Antibodies include, but are not limited to, monoclonal and chimerized monoclonal antibodies. The term "antibody" encompasses complete immunoglobulins, like monoclonal antibodies produced by recombinant expression in host cells, as well as Ang2-binding antibody fragments or "antibody-like molecules", including single-chain antibodies and linear antibodies, so-called "SMIPs" ("Small Modular Immunopharmaceuticals"), as e.g described in W002/056910. Anti- Ang2 antibody-like molecules include immunoglobulin single variable domains, as defined herein. Other examples for antibody-like molecules are immunoglobulin super family antibodies (IgSF), or CDR-grafted molecules.

[0063] "Ang2-binding molecule" refers to monovalent Ang2-binding molecules (i.e. molecules that bind to one epitope of Ang2) as well as to Ang2-binding molecules containing more than one Ang2-binding immunoglobulin single variable domain, also termed "formatted" Ang2-binding molecules. The formatted Ang-2binding molecules may, in addition to the Ang2-binding immunoglobulin single variable domains, comprise linkers and/or moieties with effector functions, e.g. half-life-extending moieties like albumin-binding immunoglobulin single variable domains, and/or a fusion partner like serum albumin and/or an attached polymer like PEG.

[0064] A formatted Ang2-binding molecule may, albeit less preferred, also comprise two identical Ang2-binding immunoglobulin single variable domains or two different immunoglobulin single variable domains that recognize the same or overlapping epitopes. In this case, the two immunoglobulin single variable domains may bind to the same or an overlapping epitope in each of the two monomers that form the Ang2 dimer. Exeprimental data including competitive ELISA assay discloses a significant improvement in the potency of the formatted Ang2 dimers when compared to the individual building blocks of mono Ang2-binding molecules (data not shown).

[0065] Specific binding of an antigen-binding protein to an antigen or epitope can be determined in any suitable manner known per se, including, for example, the assays described herein, Scatchard analysis and/or competitive binding assays, such as radioimmunoassays (RIA), enzyme immunoassays (EIA) and sandwich competition assays, and the different variants thereof known per se in the art.

[0066] Amino acid residues will be indicated according to the standard three-letter or one-letter amino acid code, as generally known and agreed upon in the art. When comparing two amino acid sequences, the term "amino acid difference" refers to insertions, deletions or substitutions of the indicated number of amino acid residues at a position of the reference sequence, compared to a second sequence. In case of substitution(s), such substitution(s) will preferably be conservative amino acid substitution(s), which means that an amino acid residue is replaced with another amino acid residue of similar chemical structure and which has little or essentially no influence on the function, activity or other biological properties of the polypeptide. Such conservative amino acid substitutions are well known in the art, for example from WO98/49185, wherein conservative amino acid substitutions preferably are substitutions in which one amino acid within the following groups (i) - (v) is substituted by another amino acid residue within the same group: (i) small aliphatic, nonpolar or slightly polar residues: Ala, Ser, Thr, Pro and Gly; (ii) polar, negatively charged residues and their (uncharged) amides: Asp, Asn, Glu and Gin; (iii) polar, positively charged residues: His, Arg and Lys; (iv) large aliphatic, nonpolar residues: Met, Leu, lie, Val and Cys; and (v) aromatic residues: Phe, Tyr and Trp. Particularly preferred conservative amino acid substitutions are as follows:

Ala into Gly or into Ser; Arg into Lys; Asn into Gin or into His; Asp into Glu; Cys into Ser; Gin into Asn; Glu into Asp; Gly into Ala or into Pro; His into Asn or into Gin; lie into Leu or into Val; Leu into lie or into Val; Lys into Arg, into Gin or into Glu; Met into Leu, into Tyr or into lie; Phe into Met, into Leu or into Tyr; Ser into Thr; Thr into Ser;Trp into Tyr; Tyr into Trp or into Phe; Val into lie or into Leu.

[0067] A polypeptide or nucleic acid molecule is considered to be "(in) essentially isolated (form)" - for example, when compared to its native biological source and/or the reaction medium or cultivation medium from which it has been obtained - when it has been separated from at least one other component with which it is usually associated in said source or medium, such as another protein/polypeptide, another nucleic acid, another biological component or macromolecule or at least one contaminant, impurity or minor component. In particular, a polypeptide or nucleic acid molecule is considered "essentially isolated" when it has been purified at least 2-fold, in particular at least 10- fold, more in particular at least 100-fold, and up to 1000-fold or more. A polypeptide or nucleic acid molecule that is "in essentially isolated form" is preferably essentially homogeneous, as determined using a suitable technique, such as a suitable chromatographical technique, such as polyacrylamide gel electrophoresis.

[0068] The term "N-terminus" (also known as the amino-terminus, NH2-terminus, N-terminal end or amine-terminus) refers to the start of a protein/polypeptide (i.e. Ang2-binding molecule) terminated by an amino acid with a free amine group (-NH2). The convention for writing peptide sequences is to put the N-terminus on the left and write the sequence from N- to C-terminus. When the protein is translated from messenger RNA, it is created from N-terminus to C-terminus.

[0069] "Sequence identity" between two Ang2-binding molecule sequences indicates the percentage of amino acids that are identical between the sequences. It may be calculated or determined as described in paragraph f) on pages 49 and 50 of W008/020079. "Sequence similarity" indicates the percentage of amino acids that either is identical or that represent conservative amino acid substitutions.

[0070] Alternative methods for numbering the amino acid residues of Vh domains, which methods can also be applied in an analogous manner to VHH domains, are known in the art. However, in the present description, claims and figures, the numbering according to Kabat and applied to VHH domains as described above will be followed, unless indicated otherwise.

[0071] An "affinity-matured" Ang2-binding molecule, in particular a VHH or a domain antibody, has one or more alterations in one or more CDRs which result in an improved affinity for Ang2, as compared to the respective parent Ang2-binding molecule. Afffinity-matured Ang2-binding molecules of the invention may be prepared by methods known in the art, for example, as described by Marks et al., 1992, Biotechnology 10:779-783, or Barbas, et al., 1994, Proc. Nat. Acad. Sci, USA 91: 3809-3813.; Shier et al., 1995, Gene 169:147-155; Yelton et al., 1995, Immunol. 155: 1994-2004; Jackson et al., 1995, J. Immunol. 154(7):3310-9; and Hawkins et al., 1992, J. Mol. Biol. 226(3): 889 896; KS Johnson and RE Hawkins, "Affinity maturation of antibodies using phage display", Oxford University Press 1996.

[0072] An "amino acid sequences of SEQ ID NO: x" means an amino acid sequence that is 100% identical with the sequence shown in the respective SEQ ID NO: x.

[0073] The terms "cancer" and "cancerous" refer to or describe the physiological condition in mammals that is typically characterized by unregulated cell growth/proliferation. Examples of cancer to be treated with an Ang2-binding molecule of the invention include but are not limited to carcinoma, lymphoma, blastoma, sarcoma, and leukemia. More particular examples of such cancers, include squamous cell cancer, small-cell lung cancer, non-small cell lung cancer, adenocarcinoma of the lung, squamous carcinoma of the lung, cancer of the peritoneum, hepatocellular cancer, gastrointestinal cancer, pancreatic cancer, glioblastoma, cervical cancer, ovarian cancer, liver cancer, bladder cancer, hepatoma, breast cancer, colon cancer, colorectal cancer, renal cell carcinoma, endometrial or uterine carcinoma, salivary gland carcinoma, kidney cancer, liver cancer, prostate cancer, vulval cancer, thyroid cancer, hepatic carcinoma, gastric cancer, melanoma, and various types of head and neck cancer. Dysregulation of angiogenesis can lead to many disorders that can be treated by compositions and methods of the invention. These disorders include both non-neoplastic and neoplastic conditions. Neoplasties include but are not limited those described above.

[0074] Non-neoplastic disorders include, but are not limited to undesired or aberrant hypertrophy, arthritis, rheumatoid arthritis (RA), psoriasis, psoriatic plaques, sarcoidosis, atherosclerosis, atherosclerotic plaques, diabetic and other proliferative retinopathies including retinopathy of prematurity, retrolental fibroplasia, neovascular glaucoma, age-related macular degeneration, diabetic macular edema, corneal neovascularization, corneal graft neovascularization, corneal graft rejection, retinal/choroidal neovascularization, neovascularization of the angle (rubeosis), ocular neovascular disease, vascular restenosis, arteriovenous malformations (AVM), meningioma, hemangioma, angiofibroma, thyroid hyperplasias (including Grave's disease), corneal and other tissue transplantation, chronic inflammation, lung inflammation, acute lung injury/ ARDS, sepsis, primary pulmonary hypertension, malignant pulmonary effusions, cerebral edema (e.g., associated with acute stroke/ closed head injury/ trauma), synovial inflammation, pannus formation in RA, myositis ossificans, hypertropic bone formation, osteoarthritis (OA), refractory ascites, polycystic ovarian disease, endometriosis, 3rd spacing of fluid diseases (pancreatitis, compartment syndrome, burns, bowel disease), uterine fibroids, premature labor, chronic inflammation such as IBD (Crohn's disease and ulcerative colitis), renal allograft rejection, inflammatory bowel disease, nephrotic syndrome, undesired or aberrant tissue mass growth (non-cancer), hemophilic joints, hypertrophic scars, inhibition of hair growth, Osier-Weber syndrome, pyogenic granuloma retrolental fibroplasias, scleroderma, trachoma, vascular adhesions, synovitis, dermatitis, preeclampsia, ascites, pericardial effusion (such as that associated with pericarditis), and pleural effusion.

[0075] The term "eye diseases" refers to proliferative retinopathies including retinopathy of prematurity, retrolental fibroplasia, neovascular glaucoma, age-related macular degeneration, diabetic macular edema, corneal neovascularization, corneal graft neovascularization, corneal graft rejection, retinal/choroidal neovascularization.

[0076] The term "chronic kidney diseases" refers to diabetic nephropathy, postrenal failure, prerenal azotemia and intrinsic renal failure.

DETAILED DESCRIPTION OF THE INVENTION

[0077] In a first aspect, the present invention relates to an Ang2-binding molecule comprising an immunoglobulin single variable domain, wherein said immunoglobulin single variable domain comprises three complementarity determining regions CDR1, CDR2 and CDR3, wherein CDR1 has an amino acid sequence shown in SEQ ID NO: 168, CDR2 has an amino acid sequence shown in SEQ ID NO: 171 and CDR3 has an amino acid sequence shown in SEQ ID NO: 175.

SEQ ID NO: 168 DYAIG

SEQ ID NO: 171 AIRSSGGSTYYADSVKG

SEQ ID NO: 175 VPAGRLRYGEQWYPIYEYDA

[0078] According to preferred embodiments, the Ang2-binding molecule comprises an immunoglobulin single variable domain, wherein said immunoglobulin single variable domain is a VHH or a domain antibody.

[0079] According to a preferred embodiment, the Ang2-binding molecule comprises an immunoglobulin single variable domain, wherein said immunoglobulin single variable domain is a VHH.

[0080] According to specific embodiments, said VHH consists of an immunoglobulin single variable domain having a sequence of SEQ ID NO: 166.

[0081] In another aspect, the present invention relates to an Ang2-binding molecule consisting of said immunoglobulin single variable domain.

[0082] The sequence of the Ang2 binders according to the invention can be modified at their N-terminus (i.e. deletion or exchange of the first amino acid) without significant reduction of their binding activity. This modification enhances the co-/post-translational cleavage of N-terminal methionine during intracellular/cytoplasmic expression in bacterial hosts (e.g. but not limited to Escherichia coli).

[0083] In one aspect said VHH consisting of an immunoglobulin single variable domain has a modification or exchange on N terminus, wherein said modification is a deletion of a first amino acid and said exchange is a replacement of the first amino acid by another amino acid.

[0084] In one of preferred embodiments the first amino acid on N terminus is Valine (V) or Aspartic acid (D) replaced by e.g. by Alanine (A).

[0085] Ang2-binding components with improved properties in view of therapeutic application, e.g. enhanced affinity or decreased immunogenicity, may be obtained from individual Ang2-binding components of the invention by techniques such as affinity maturation (for example, starting from synthetic, random or naturally occurring immunoglobulin sequences), CDR grafting, humanizing, combining fragments derived from different immunoglobulin sequences, PCR assembly using overlapping primers, and similar techniques for engineering immunoglobulin sequences well known to the skilled person; or any suitable combination of any of the foregoing. Reference is, for example, made to standard handbooks, as well as to the further description and Examples.

[0086] In yet another embodiment, the representatives of the class of Ang2-binding immunoglobulin single variable domains of the invention which include the CDRs as set out above have amino acid sequences that correspond to the amino acid sequence of a naturally occurring VH domain that has been "camelized", i.e. by replacing one or more amino acid residues in the amino acid sequence of a naturally occurring variable heavy chain from a conventional 4-chain antibody by one or more amino acid residues that occur at the corresponding position(s) in a VHH domain of a heavy chain antibody. This can be performed in a manner known per se, which will be clear to the skilled person, and reference is additionally be made to WO 1994/04678. Such camelization may preferentially occur at amino acid positions which are present at the VH-VL interface and at the so-called Camelidae Hallmark residues (see for example also WO 1994/04678). A detained description of such "humanization" and "camelization" techniques and preferred framework region sequences consistent therewith can additionally be taken from e.g. pp. 46 and pp. 98 of WO 2006/040153 and pp. 107 of WO 2006/122786.

[0087] The Ang2-binding components of the invention, e.g. immunoglobulin single variable domains and or polypeptides containing them, have specificity for Ang2 in that they comprise one or more immunoglobulin single variable domains specifically binding to one or more epitopes within the Ang2 molecule.

[0088] Specific binding of an Ang2-binding component to its antigen Ang2 can be determined in any suitable manner known per se, including, for example, the assays described herein, Scatchard analysis and/or competitive binding assays, such as radioimmunoassays (RIA), enzyme immunoassays (EIA and ELISA) and sandwich competition assays, and the different variants thereof known per se in the art.

[0089] According to another embodiment, the immunoglobulin single variable domains are domain antibodies, as defined herein.

[0090] In such case, immunoglobulin single variable domains as defined above present in the monospecific binding molecules of the invention have sequences that correspond to the amino acid sequence of a naturally occurring VH domain that has been "camelized", i.e. by replacing one or more amino acid residues in the amino acid sequence of a naturally occurring variable heavy chain from a conventional 4-chain antibody by one or more amino acid residues that occur at the corresponding position(s) in a VHH domain of a heavy chain antibody. This can be performed in a manner known per se, which will be clear to the skilled person, and reference is additionally be made to WO 94/04678. Such camelization may preferentially occur at amino acid positions which are present at the VH-VL interface and at the so-called Camelidae Hallmark residues (see for example also WO 94/04678). A detained description of such "humanization" and "camelization" techniques and preferred framework region sequences consistent therewith can additionally be taken from e.g. pp. 46 and pp. 98 of WO 2006/040153 and pp. 107 of WO 2006/122786.

[0091] The binding components have specificity for Ang2, in that they comprise one immunoglobulin single variable domain specifically binding to the Ang2 molecule.

[0092] Specific binding of a binding component to its antigen Ang2 can be determined in any suitable manner known perse, including, for example, the assays described herein, Scatchard analysis and/or competitive binding assays, such as radioimmunoassays (RIA), enzyme immunoassays (EIA and ELISA) and sandwich competition assays, and the different variants thereof known per se in the art.

[0093] The present invention also relates to a nucleic acid encoding the Ang2-binding molecule according to the invention.

[0094] In a preferred embodiment an Ang2-binding molecule according to the invention when expressed in Escherichia coli is encoded by the nucelotide seuquence: SEQ ID NO: 179 [0095]

GAAGTGCAACTGGTTGAGTCAGGTGGCGGACTGGTGCAACCGGGTGGTTCACTGCGCCTG

AGTTGCGCAGTTAGCGGTATTACCCTGGATGATTATGCAATTGGTTGGTTTCGCCAAGCCC

CAGGCAAAGAGCGTGAAGGCGTTAGCGCAATTCGTAGCAGCGGTGGTAGCACCTATTATG

CCGATTCAGTTAAAGGCCGTTTTACGATCAGCAGCGATAACAGTAAAAACACGGTTTATCTG

CAAATGAACTCATTACGTCCAGAGGACACTGCAGTTTACTATTGCGCAGCAGTTCCGGCAG

GTCGTCTGCGTTATGGTGAACAGTGGTATCCGATTTATGAATATGATGCATGGGGTCAAGG

TACACTGGTTACAGTGAGTAGC

[0096] The invention relates to nucleic acid molecules that encode Ang2 binding monospecific binding molecules of the invention. Such nucleic acid molecules will also be referred to herein as "nucleic acids of the invention" and may also be in the form of a genetic construct, as defined herein. A nucleic acid of the invention may be genomic DNA, cDNA or synthetic DNA (such as DNA with a codon usage that has been specifically adapted for expression in the intended host cell or host organism). According to one embodiment of the invention, the nucleic acid of the invention is in essentially isolated form, as defined hereabove.

[0097] Further aspect of the invention relates to an expression vector comprising the nucleic acid molecule encoding said Ang2- binding molecule according to invention.

[0098] In preferred embodiments, the nucleic acid of the invention may also be in the form of, may be present in and/or may be part of a vector, such as for example a plasmid, cosmid or YAC.

[0099] The vector may especially be an expression vector, i.e. a vector that can provide for expression of the monospecific binding molecule in vitro and/or in vivo (i.e. in a suitable host cell, host organism and/or expression system). Such expression vector generally comprises at least one nucleic acid of the invention that is operably linked to one or more suitable regulatory elements, such as promoter(s), enhancer(s), terminator(s), and the like. Such elements and their selection in view of expression of a specific sequence in a specific host are common knowledge of the skilled person. Specific examples of regulatory elements and other elements useful or necessary for expressing molecules of the invention, such as promoters, enhancers, terminators, integration factors, selection markers, leader sequences, reporter genes, and the like, are disclosed e.g. on pp. 131 to 133 of WO 2006/040153.

[0100] Such vectors express or are capable of expressing one or more monospecific binding molecules of the invention; and/or contain a nucleic acid of the invention.

[0101] The nucleic acids of the invention may be prepared or obtained in a manner known per se (e.g. by automated DNA synthesis and/or recombinant DNA technology), based on the information on the amino acid sequences for the polypeptides of the invention given herein, and/or can be isolated from a suitable natural source.

[0102] In another aspect, the invention relates to a host cell carrying one or more expression vectors comprising a nucleic acid molecule encoding the Ang2-binding molecule according to the invention.

[0103] According to a particularly preferred embodiment, said host cells are bacterial cells; other useful cells are yeast cells, fungal cells or mammalian cells.

[0104] Suitable bacterial cells include cells from gram-negative bacterial strains such as strains of Escherichia coli, Proteus, and Pseudomonas, and gram-positive bacterial strains such as strains of Bacillus, Streptomyces, Staphylococcus, and Lactococcus. Suitable fungal cell include cells from species of Trichoderma, Neurospora, and Aspergillus. Suitable yeast cells include cells from species of Saccharomyces (for example Saccharomyces cerevisiae), Schizosaccharomyces (for example Schizosaccharomyces pombe), Pichia (for example Pichia pastoris and Pichia methanolica), and Hansenula.

[0105] Suitable mammalian cells include for example CHO cells, BHK cells, HeLa cells, COS cells, and the like. However, amphibian cells, insect cells, plant cells, and any other cells used in the art for the expression of heterologous proteins can be used as well.

[0106] The invention further provides methods of manufacturing a monospecific binding molecule of the invention, such methods generally comprising the steps of: • culturing host cells comprising a nucleic acid capable of encoding a monospecific binding molecule under conditions that allow expression of the monospecific binding molecule of the invention; and • recovering or isolating the polypeptide expressed by the host cells from the culture; and • optionally further purifying and/or modifying and/or formulating the monospecific binding molecule of the invention.

[0107] The preferred embodiment represents a method for producing an Ang2 binding molecule having sequence SEQ ID NO: 166 comprising the steps of: 1. (a) transfecting a host cell with one or more said vectors 2. (b) culturing said host cell, and 3. (c) recovering and purifying said Ang2 binding molecule.

[0108] For production on an industrial scale, preferred host organisms include strains of E. coli, Pichia pastoris, and S. cerevisiae that are suitable for large scale expression, production and fermentation, and in particular for large scale pharmaceutical expression, production and fermentation.

[0109] The choice of the specific expression system depends in part on the requirement for certain post-translational modifications, more specifically glycosylation. The production of a monospecific binding molecule of the invention for which glycosylation is desired or required would necessitate the use of mammalian expression hosts that have the ability to glycosylate the expressed protein. In this respect, it will be clear to the skilled person that the glycosylation pattern obtained (i.e. the kind, number and position of residues attached) will depend on the cell or cell line that is used for the expression.

[0110] Monospecific binding molecules of the invention may be produced either in a cell as set out above intracellullarly (e.g. in the cytosol, in the periplasma or in inclusion bodies) and then isolated from the host cells and optionally further purified; or they can be produced extracellularly (e.g. in the medium in which the host cells are cultured) and then isolated from the culture medium and optionally further purified.

[0111] Methods and reagents used for the recombinant production of polypeptides, such as specific suitable expression vectors, transformation or transfection methods, selection markers, methods of induction of protein expression, culture conditions, and the like, are known in the art. Similarly, protein isolation and purification techniques useful in a method of manufacture of a polypeptide of the invention are well known to the skilled person.

[0112] The invention further relates to a product or composition containing or comprising at least one Ang2 binding monospecific binding molecule of the invention and optionally one or more further components of such compositions known perse, i.e. depending on the intended use of the composition.

[0113] In further aspect, the invention relates to use of an Ang2 binding monospecific binding molecule of the invention as a medicament.

[0114] In still another aspect, the invention relates to use of an Ang2 binding monospecific binding molecule of the invention for method of treating of cancer, cancerous or eye diseases.

[0115] For pharmaceutical use, an Ang2 binding monospecific binding molecule of the invention or a polypeptide containing same may be formulated as a pharmaceutical preparation or composition comprising at least one such monospecific binding molecule of the invention and at least one physiologically acceptable carrier, diluent or excipient and/or adjuvant, and optionally one or more further pharmaceutically active polypeptides and/or compounds. By means of non-limiting examples, such a formulation may be in a form suitable for oral administration, for parenteral administration (such as by intravenous, intramuscular or subcutaneous injection or intravenous infusion), for topical administration, for administration by inhalation, by a skin patch, by an implant, by a suppository, etc. Such suitable administration forms - which may be solid, semi-solid or liquid, depending on the manner of administration - as well as methods and carriers for use in the preparation thereof, will be clear to the skilled person, and are further described herein.

[0116] Thus, in a further aspect, the invention relates to a pharmaceutical composition that contains at least one Ang2 binding monospecific binding molecule, in particular one immunoglobulin single variable domain of the invention as defined above or a polypeptide containing same and at least one suitable carrier, diluent or excipient (i.e. suitable for pharmaceutical use), and optionally one or more further active substances.

[0117] The Ang2 binding monospecific binding molecules of the invention may be formulated and administered in any suitable manner known per se: Reference, in particular for the immunoglobulin single variable domains, is for example made to WO 2004/041862, WO 2004/041863, WO 2004/041865, WO 2004/041867 and WO 2008/020079, as well as to the standard handbooks, such as Remington's Pharmaceutical Sciences, 18th Ed., Mack Publishing Company, USA (1990), Remington, the Science and Practice of Pharmacy, 21th Edition, Lippincott Williams and Wilkins (2005); or the Handbook of Therapeutic Antibodies (S. Dubel, Ed.), Wiley, Weinheim, 2007 (see for example pages 252-255).

[0118] For example, an immunoglobulin single variable domain of the invention as defined above may be formulated and administered in any manner known per se for conventional antibodies and antibody fragments (including ScFv's and diabodies) and other pharmaceutically active proteins. Such formulations and methods for preparing the same will be clear to the skilled person, and for example include preparations suitable for parenteral administration (for example intravenous, intraperitoneal, subcutaneous, intramuscular, intraluminal, intra-arterial or intrathecal administration) or for topical (i.e. transdermal or intradermal) administration.

[0119] Preparations for parenteral administration may for example be sterile solutions, suspensions, dispersions or emulsions that are suitable for infusion or injection. Suitable carriers or diluents for such preparations for example include, without limitation, sterile water and pharmaceutically acceptable aqueous buffers and solutions such as physiological phosphate-buffered saline, Ringer's solutions, dextrose solution, and Hank's solution; water oils; glycerol; ethanol; glycols such as propylene glycol or as well as mineral oils, animal oils and vegetable oils, for example peanut oil, soybean oil, as well as suitable mixtures thereof. Usually, aqueous solutions or suspensions will be preferred.

[0120] Thus, the Ang2 binding monospecific binding molecule of the invention may be systemically administered, e.g., orally, in combination with a pharmaceutically acceptable vehicle such as an inert diluent or an assimilable edible carrier. For oral therapeutic administration, the molecule of the invention may be combined with one or more excipients and used in the form of ingestible tablets, buccal tablets, troches, capsules, elixirs, suspensions, syrups, wafers, and the like. Such compositions and preparations should contain at least 0.1% of the Ang2-binding molecule of the invention. Their percentage in the compositions and preparations may, of course, be varied and may conveniently be between about 2 to about 60% of the weight of a given unit dosage form. The amount of the molecule of the invention in such therapeutically useful compositions is such that an effective dosage level will be obtained.

[0121] The tablets, pills, capsules, and the like may also contain binders, excipients, disintegrating agents, lubricants and sweetening or flavouring agents, for example those mentioned on pages 143-144 of WO 08/020079. When the unit dosage form is a capsule, it may contain, in addition to materials of the above type, a liquid carrier, such as a vegetable oil or a polyethylene glycol. Various other materials may be present as coatings or to otherwise modify the physical form of the solid unit dosage form. For instance, tablets, pills, or capsules may be coated with gelatin, wax, shellac or sugar and the like. A syrup or elixir may contain the molecules of the invention, sucrose or fructose as a sweetening agent, methyl and propylparabens as preservatives, a dye and flavoring such as cherry or orange flavor. Of course, any material used in preparing any unit dosage form should be pharmaceutically acceptable and substantially non-toxic in the amounts employed. In addition, the molecules of the invention may be incorporated into sustained-release preparations and devices.

[0122] Preparations and formulations for oral administration may also be provided with an enteric coating that will allow the constructs of the invention to resist the gastric environment and pass into the intestines. More generally, preparations and formulations for oral administration may be suitably formulated for delivery into any desired part of the gastrointestinal tract. In addition, suitable suppositories may be used for delivery into the gastrointestinal tract.

[0123] The Ang2 binding monospecific binding molecules of the invention may also be administered intravenously or intraperitoneally by infusion or injection, as further described on pages 144 and 145 of WO 2008/020079.

[0124] For topical administration of the Ang2 binding monospecific binding molecules of the invention, it will generally be desirable to administer them to the skin as compositions or formulations, in combination with a dermatologically acceptable carrier, which may be a solid or a liquid, as further described on page 145 of WO 2008/020079.

[0125] Generally, the concentration of the Ang2 binding monospecific binding molecules of the invention in a liquid composition, such as a lotion, will be from about 0.1-25 wt-%, preferably from about 0.5-10 wt-%. The concentration in a semi-solid or solid composition such as a gel or a powder will be about 0.1-5 wt-%, preferably about 0.5-2.5 wt-%.

[0126] The amount of the Ang2 binding monospecific binding molecules of the invention required for use in treatment will vary not only with the particular monospecific binding molecule selected, but also with the route of administration, the nature of the condition being treated and the age and condition of the patient and will be ultimately at the discretion of the attendant physician or clinician. Also, the dosage of the monospecific binding molecules of the invention varies depending on the target cell, tumor, tissue, graft, or organ. The desired dose may conveniently be presented in a single dose or as divided doses administered at appropriate intervals, for example, as two, three, four or more sub-doses per day. The sub-dose itself may be further divided, e.g., into a number of discrete loosely spaced administrations; such as multiple inhalations from an insufflator or by application of a plurality of drops into the eye.

[0127] An administration regimen may include long-term, daily treatment. By "long-term" is meant at least two weeks and preferably, several weeks, months, or years of duration. Necessary modifications in this dosage range may be determined by one of ordinary skill in the art using only routine experimentation given the teachings herein. See Remington's Pharmaceutical Sciences (Martin, E.W., ed. 4), Mack Publishing Co., Easton, PA. The dosage can also be adjusted by the individual physician in the event of any complication.

[0128] According to a further embodiment, the invention relates to the use of Ang2 binding monospecific binding molecules of the invention, i.e. immunoglobulin single variable domains or polypeptides containing them, for therapeutic purposes, such as • for the prevention, treatment and/or alleviation of a disorder, disease or condition, especially in a human being, that is associated with Ang2-mediated and/or Ang2-related effects on angiogenesis or that can be prevented, treated or alleviated by modulating the Notch signaling pathway and/or the Tie2 signalling pathway with a monospecific binding molecule according to the invention; • in a method of treatment of a patient in need of such therapy, such method comprising administering, to a subject in need thereof, a pharmaceutically active amount of at least one monospecific binding molecule of the invention, e.g. an immunoglobulin single variable domain, or a pharmaceutical composition containing same; • for the preparation of a medicament for the prevention, treatment or alleviation of disorders, diseases or conditions associated with Ang2-mediated and/or Ang2-mediated effects on angiogenesis; • as an active ingredient in a pharmaceutical composition or medicament used for the above purposes.

[0129] According to a specific aspect, said disorder disorder, disease or condition is a cancer or cancerous disease, as defined herein.

[0130] In preferred embodiments, the invention relates to said pharmaceutical composition for treatment of cancer and cancerous diseases, such as breast, renal cell carcinoma, ovarian cancer and pancreatic cancer.

[0131] According to another aspect, the disease is an eye disease associated with Ang2-mediated and/or Ang2-mediated effects on angiogenesis or which can be treated or alleviated by modulating the Notch signaling pathway and/or the Tie2 signalling pathway with an Ang2 binding monospecific binding molecule of the invention.

[0132] In another preferred embodiments, the invention relates to said pharmaceutical composition for treatment of eye diseases, such as age-related macular degeneration and diabetic retinopathy.

[0133] In still another preferred embodiments, the invention relates to said pharmaceutical composition for treatment of chronic kidney diseases.

[0134] Depending on cancer/ cancerous diseases, eye diseases and/or chronic kidney diseases to be treated, an Ang2 binding monospecific binding molecule of the invention may be used on its own or in combination with one or more additional therapeutic agents.

[0135] In preferred embodiments, the invention relates to the pharmaceutical composition comprising, as the active ingredient one or more said Ang-2 binding molecules of the invention, further comprising one or more additional therapeutic agents, such as chemotherapeutic agents like DNA damaging agents and/or anti-mitotic drugs in cancer cells (e.g. taxol) or therapeutically active compounds that inhibit angiogenesis (an anti- angiogenic drug such as anti VEGF/VEGF receptor inhibitor, e.g. avastin,nitedanib and sunitinib), or signal transduction pathway inhibitors such as mTOR inhibitors (e.g. temsirolimus) or hormonal therapy agents (e.g. tamoxifen).

[0136] The additional therapeutic agent may be administered simultaneously with, optionally as a component of the same pharmaceutical preparation, or before or after administration of the monospecific binding molecule.

[0137] In certain embodiments, the additional therapeutic agent may be, without limitation (and in the case of the receptors, including the respective ligands), one or more inhibitors selected from the group of inhibitors of EGFR, VEGF, VEGFR, HER2-neu, Her3, AuroraA, AuroraB, PLK and PI3 kinase, FGFR, PDGFR, Raf, KSP, PDK1, PTK2, IGF-Ror IR.

[0138] Further examples of additional therapeutic agents are inhibitors of CDK, Akt, src/bcr abl, cKit, cMet/HGF, c-Myc, Flt3, HSP90, hedgehog antagonists, inhibitors of JAK/STAT, Mek, mTor, NFkappaB, the proteasome, Rho, an inhibitor of wnt signaling or an inhibitor of the ubiquitination pathway or another inhibitor of the Notch signaling pathway.

[0139] Examples for Aurora inhibitors are, without limitation, PHA-739358, AZD-1152, AT 9283, CYC-116, R-763, VX-680, VX-667, MLN-8045, PF-3814735.

[0140] An example for a PLK inhibitor is GSK-461364.

[0141] Examples for VEGF inhibitor are avastin (Roche), aflibercept (Regeneron,) [0142] Examples for raf inhibitors are BAY-73-4506 (also a VEGFR inhibitor), PLX 4032, RAF-265 (also in addition a VEGFR inhibitor), sorafenib (also in addition a VEGFR inhibitor), and XL 281.

[0143] Examples for KSP inhibitors are ispinesib, ARRY-520, AZD-4877, CK-1122697, GSK 246053A, GSK-923295, MK-0731, and SB-743921.

[0144] Examples for a src and/or bcr-abl inhibitors are dasatinib, AZD-0530, bosutinib, XL 228 (also an IGF-1R inhibitor), nilotinib (also a PDGFR and cKit inhibitor), imatinib (also a cKit inhibitor), and NS-187.

[0145] An example for a PDK1 inhibitor is BX-517.

[0146] An example for a Rho inhibitor is BA-210.

[0147] Examples for PI3 kinase inhibitors are PX-866, BEZ-235 (also an mTor inhibitor), XL 418 (also an Akt inhibitor), XL-147, and XL 765 (also an mTor inhibitor).

[0148] Examples for inhibitors of cMet or HGF are XL-184 (also an inhibitor of VEGFR, cKit, Flt3), PF-2341066, MK-2461, XL-880 (also an inhibitor of VEGFR), MGCD-265 (also an inhibitor of VEGFR, Ron, Tie2), SU-11274, PHA-665752, AMG-102, and AV-299.

[0149] An example for a c-Myc inhibitor is CX-3543.

[0150] Examples for Flt3 inhibitors are AC-220 (also an inhibitor of cKit and PDGFR), KW 2449, lestaurtinib (also an inhibitor of VEGFR, PDGFR, PKC), TG-101348 (also an inhibitor of JAK2), XL-999 (also an inhibitor of cKit, FGFR, PDGFR and VEGFR), sunitinib (also an inhibitor of PDGFR, VEGFR and cKit), and tandutinib (also an inhibitor of PDGFR, and cKit).

[0151] Examples for HSP90 inhibitors are tanespimycin, alvespimycin, IPI-504 and CNF 2024.

[0152] Examples for JAK/STAT inhibitors are CYT-997 (also interacting with tubulin), TG 101348 (also an inhibitor of Flt3), and XL-019.

[0153] Examples for Mek inhibitors are ARRY-142886, PD-325901, AZD-8330, and XL 518.

[0154] Examples for mTor inhibitors are temsirolimus, AP-23573 (which also acts as a VEGF inhibitor), everolimus (a VEGF inhibitor in addition). XL-765 (also a PI3 kinase inhibitor), and BEZ-235 (also a PI3 kinase inhibitor).

[0155] Examples for Akt inhibitors are perifosine, GSK-690693, RX-0201, and triciribine.

[0156] Examples for cKit inhibitors are AB-1010, OSI-930 (also acts as a VEGFR inhibitor), AC-220 (also an inhibitor of Flt3 and PDGFR), tandutinib (also an inhibitor of Flt3 and PDGFR), axitinib (also an inhibitor of VEGFR and PDGFR), XL-999 (also an inhibitor of Flt3, PDGFR, VEGFR, FGFR), sunitinib (also an inhibitor of Flt3, PDGFR, VEGFR), and XL-820 (also acts as a VEGFR- and PDGFR inhibitor), imatinib (also a bcr-abl inhibitor), nilotinib (also an inhibitor of bcr-abl and PDGFR).

[0157] Examples for hedgehog antagonists are IPI-609 and CUR-61414.

[0158] Examples for CDK inhibitors are seliciclib, AT-7519, P-276, ZK-CDK (also inhibiting VEGFR2 and PDGFR), PD-332991, R-547, SNS-032, PHA-690509, and AG 024322.

[0159] Examples for proteasome inhibitors are bortezomib, carfilzomib, and NPI-0052 (also an inhibitor of NFkappaB).

[0160] An example for an NFkappaB pathway inhibitor is NPI-0052.

[0161] An example for an ubiquitination pathway inhibitor is HBX-41108.

[0162] In preferred embodiments, the additional therapeutic agent is an anti-angiogenic agent.

[0163] Examples for anti-angiogenic agents are inhibitors of the FGFR, PDGFR and VEGFR or the respective ligands (e.g VEGF inhibitors like pegaptanib or the anti-VEGF antibody bevacizumab), and thalidomides, such agents being selected from, without limitation, bevacizumab, motesanib, CDP-791, SU-14813, telatinib, KRN-951, ZK-CDK (also an inhibitor of CDK), ABT-869, BMS-690514, RAF-265, IMC-KDR, IMC-18F1, IMiDs (immunomodulatory drugs), thalidomide derivative CC-4047, lenalidomide, ENMD 0995, IMC-D11, Ki 23057, brivanib, cediranib, XL-999 (also an inhibitor of cKit and Flt3), 1B3, CP 868596, IMC 3G3, R-1530 (also an inhibitor of Flt3), sunitinib (also an inhibitor of cKit and Flt3), axitinib (also an inhibitor of cKit), vemurafenib (also known as PLX4032, RG7204 or RO5185426, marketed as zelboraf) a B-Raf enzyme inhibitor, crizotinib known as an ALK (anaplastic lymphoma kinase) and ROS1 (c-ros oncogenel, receptor tyrosine kinase) inhibitor, lestaurtinib (also an inhibitor of Flt3 and PKC), vatalanib, tandutinib (also an inhibitor of Flt3 and cKit), pazopanib, GW 786034, PF-337210, IMC-1121B, AVE-0005, AG-13736, E-7080, CHIR 258, sorafenib tosylate (also an inhibitor of Raf), RAF-265 (also an inhibitor of Raf), vandetanib, CP-547632, OSI-930, AEE-788 (also an inhibitor of EGFR and Her2), BAY-57-9352 (also an inhibitor of Raf), BAY-73-4506 (also an inhibitor of Raf), XL 880 (also an inhibitor of cMet), XL-647 (also an inhibitor of EGFR and EphB4), XL 820 (also an inhibitor of cKit), and nilotinib (also an inhibitor of cKit and brc-abl) and nitedanib.

[0164] The additional therapeutic agent may also be selected from EGFR inhibitors; it may be a small molecule EGFR inhibitor or an anti-EGFR antibody. Examples for anti-EGFR antibodies, without limitation, are cetuximab, panitumumab, matuzumab; an example for a small molecule EGFR inhibitor is gefitinib. Another example for an EGFR modulator is the EGF fusion toxin.

[0165] Among the EGFR and Her2 inhibitors useful for combination with the molecule of the invention are lapatinib, gefitinib, erlotinib, cetuximab, trastuzumab, nimotuzumab, zalutumumab, vandetanib (also an inhibitor of VEGFR), pertuzumab, XL-647, HKI-272, BMS-599626 ARRY-334543, AV 412, mAB-806, BMS-690514, JNJ-26483327, AEE-788 (also an inhibitor of VEGFR), ARRY-333786, IMC-11F8, Zemab.

[0166] Other agents that may be advantageously combined in a therapy with the Ang2 binding monospecific binding molecule of the invention are tositumumab and ibritumomab tiuxetan (two radiolabelled anti-CD20 antibodies), alemtuzumab (an anti-CD52 antibody), denosumab, (an osteoclast differentiation factor ligand inhibitor), galiximab (a CD80 antagonist), ofatumumab (a CD20 inhibitor), zanolimumab (a CD4 antagonist), SGN40 (a CD40 ligand receptor modulator), rituximab (a CD20 inhibitor) or mapatumumab (a TRAIL-1 receptor agonist) or OMP-21M18 (DII4 inhibitors).

[0167] Other chemotherapeutic drugs that may be used in combination with the molecules of the present invention are selected from, but not limited to hormones, hormonal analogues and antihormonals (e.g. tamoxifen, toremifene, raloxifene, fulvestrant, megestrol acetate, flutamide, nilutamide, bicalutamide, cyproterone acetate, finasteride, buserelin acetate, fludrocortisone, fluoxymesterone, medroxyprogesterone, octreotide, arzoxifene, pasireotide, vapreotide), aromatase inhibitors (e.g. anastrozole, letrozole, liarozole, exemestane, atamestane, formestane), LHRH agonists and antagonists (e.g. goserelin acetate, leuprolide, abarelix, cetrorelix, deslorelin, histrelin, triptorelin), antimetabolites (e.g. antifolates like methotrexate, pemetrexed, pyrimidine analogues like 5 fluorouracil, capecitabine, decitabine, nelarabine, and gemcitabine, purine and adenosine analogues such as mercaptopurine thioguanine, cladribine and pentostatin, cytarabine, fludarabine); antitumor antibiotics (e.g. anthracyclines like doxorubicin, daunorubicin, epirubicin and idarubicin, mitomycin-C, bleomycin dactinomycin, plicamycin, mitoxantrone, pixantrone, streptozocin); platinum derivatives (e.g. cisplatin, oxaliplatin, carboplatin, lobaplatin, satraplatin); alkylating agents (e.g. estramustine, meclorethamine, melphalan, chlorambucil, busulphan, dacarbazine, cyclophosphamide, ifosfamide, hydroxyurea, temozolomide, nitrosoureas such as carmustine and lomustine, thiotepa); antimitotic agents (e.g. vinca alkaloids like vinblastine, vindesine, vinorelbine, vinflunine and vincristine; and taxanes like paclitaxel, docetaxel and their formulations, larotaxel; simotaxel, and epothilones like ixabepilone, patupilone, ZK-EPO); topoisomerase inhibitors (e.g. epipodophyllotoxins like etoposide and etopophos, teniposide, amsacrine, topotecan, irinotecan) and miscellaneous chemotherapeutics such as amifostine, anagrelide, interferone alpha, procarbazine, mitotane, and porfimer, bexarotene, celecoxib.

[0168] Particularly preferred combination partners of the molecules of the present invention are VEGF antagonists, like bevacizumab (Avastin®), nitedanib, Sorafenib and Sunitinib.

[0169] According to another embodiment of the invention, there is provided a method of diagnosing a disease by 1. a) contacting a sample with an Ang2 binding molecule of the invention as defined above, and 2. b) detecting binding of said binding molecule to said sample, and 3. c) comparing the binding detected in step (b) with a standard, wherein a difference in binding relative to said sample is diagnostic of a disease or disorder associated with VEGF- and/or Ang2-mediated effects on angiogenesis.

[0170] For this and other uses, it may be useful to further modify an Ang2 binding monospecific binding molecule of the invention, such as by introduction of a functional group that is one part of a specific binding pair, such as the biotin-(strept)avidin binding pair. Such a functional group may be used to link the binding molecule of the invention to another protein, polypeptide or chemical compound that is bound to the other half of the binding pair, i.e. through formation of the binding pair. For example, such monospecific binding molecule of the invention may be conjugated to biotin, and linked to another protein, polypeptide, compound or carrier conjugated to avidin or streptavidin. For example, such a conjugated monospecific binding molecule of the invention may be used as a reporter, for example in a diagnostic system where a detectable signal-producing agent is conjugated to avidin or streptavidin.

[0171] The efficacy of Ang2 binding monospecific binding molecule of the invention or polypeptides, and of compositions comprising the same, can be tested using any suitable in vitro assay, cell-based assay, in vivo assay and/or animal model known per se, or any combination thereof, depending on the specific disease or disorder of interest. Suitable assays and animal models will be clear to the skilled person, and for example include the assays described herein and used in the Examples below, e.g. a proliferation assay.

[0172] Ang2 binding monospecific binding molecules of the invention have undergone an extensive sequence optimization process involving affinity maturation, humanization and removal of potential posttranslational modification sites to ensure low immunogenicity potential in man and improved biophysical stability. Unexpectedly, the data show that such monospecific binding molecules of the invention have properties that are superior to those of binding molecules of the prior art. Among such properties are high selectivity for Ang2 neutralization as compared to Ang1 neutralization, as can e.g. be taken from the data of Figures 9 to 10, 13 to 14, 16 to 19; complete inhibition of the Ang2-Tie2 interaction with high potency, as can e.g. be taken from the ELISA data of Figures 6, 9, 13, 16, 18 and 20 and Tables 12 to 13, 16 to 17, 20 to 22, as well as the IC50 (nM) values for VHHs in the AlphaScreen assay as shown e.g. in Table 7 (Example 7); and the affinity KD (nM) of purified VHHs on recombinant human Ang2, cyno Ang2, mouse Ang2 in Tables 8, 14, 18 and 23.

[0173] This indicates that Ang2 binding monospecific binding molecules of the invention are promising candidates to have therapeutic efficacy in diseases and disorders associated with Ang2-mediated effects on angiogenesis, such as cancer, cancerous diseases, eye diseases and/or chronic kidney diseases.

Brief description of the Figures: [0174] The axes annotation in Figures 1 to 4, 6 to 7, 9 to 10, 12 to 14 and 16 to 20: X axes stand for OD 450 (nm) and Y axes stand for log competitor (M).

Figure 1 (Figure 1-1Ato 1-2C): Purified VHHs blocking hAng2-hTie2 interaction (ELISA).

Figure 2 (Figure 2-1A to 2-2C): Purified VHHs blocking mAng2-mTie2 interaction (ELISA).

Figure 3 (Figure 3Ato 3B): Purified VHHs blocking cAng2-cTie2 interaction (ELISA).

Figure 4 (Figure 4Ato 4I): Purified VHHs blocking hAng1-hTie2 interaction (ELISA).

Figure 5: Sequence alignment of affinity matured variants of VHH 28D10. The amino acid sequence is aligned to the human germline VH3/JH consensus sequence. Residues are numbered according to Kabat, CDRs are shown in bold according to AbM definition. Residues that have been substituted are underlined.

Figure 6 (Figure 6A to 6C): Purified affinity matured variants of VHH 28D10 blocking hAng2-hTie2 interaction (ELISA).

Figure 7 (Figure 7A to 7C): Purified affinity matured variants of VHH 28D10 blocking hAngl-hTie2 interaction (ELISA).

Figure 8 (Figure 8A to 8B): Sequence alignment of VHH 1D01 with hVH3-JH consensus (A) and of sequence optimized variants of VHH 1D01 (B). The amino acid sequence is aligned to the human germline VH3/JH consensus sequence. Residues are numbered according to Kabat, CDRs are shown in bold according to AbM definition. Residues to be mutated to their human counterpart are underlined. Potential post-translational modification sites to be tackled are boxed.

Figure 9 (Figure 9-1A to 9-3B): Purified sequence optimized variants of VHH 1D01 blocking hAng2-hTie2 (10-1), mAng2-mTie2 (10-2) and cAng2-cTie2 (11-3) interaction (ELISA).

Figure 10: Purified sequence optimized variants of VHH 1D01 blocking hAng1-hTie2 interaction (ELISA).

Figure 11 (Figure 11Ato 11C): Sequence alignment of VHH 37F02 with hVH3-JH consensus (A), of cycle 1 (B) and of cycle 2 (C) sequence optimized variants of VHH 37F02. The amino acid sequence is aligned to the human germline VH3/JH consensus sequence. Residues are numbered according to Kabat, CDRs are shown in bold according to AbM definition. Residues to be mutated to their human counterpart are underlined. Potential post-translational modification sites to be tackled are boxed.

Figure 12 (Figure 12-1 to 12-3): Purified cycle 1 sequence optimized variants of VHH 37F02 blocking hAng2-hTie2 (14-1), mAng2-mTie2 (14-2) and cAng2-cTie2 (14-3) interaction (ELISA).

Figure 13 (Figure 13-1Ato 13-3): Purified cycle 2 sequence optimized variants of VHH 37F02 blocking hAng2-hTie2 (15-1), mAng2-mTie2 (15-2) and cAng2-cTie2 (15-3) interaction (ELISA).

Figure 14: Purified cycle 2 sequence optimized variants of VHH 37F02 blocking hAng1-hTie2 interaction (ELISA).

Figure 15 (Figure 15A to 15D): Sequence alignment of VHH 28D10 with hVH3-JH consensus (A), of cycle 1 sequence optimized variants (B), of cycle 2 variants (C) and of cycle 3 (D) sequence optimized variants of VHH 28D10. The amino acid sequence is aligned to the human germline VH3/JH consensus sequence. Residues are numbered according to Kabat, CDRs are shown in bold according to AbM definition. Residues to be mutated to their human counterpart are underlined. Potential post-translational modification sites to be tackled are boxed.

Figure 16 (Figure 16-1A to 16-3C): Purified cycle 1 sequence optimized variants of VHH 28D10 blocking hAng2-hTie2 (18-1), mAng2-mTie2 (18-2) and cAng2-cTie2 (18-3) interaction (ELISA).

Figure 17 (Figure 17A to 17B): Purified cycle 1 sequence optimized variants of VHH 28D10 blocking hAng1-hTie2 interaction (ELISA).

Figure 18 (Figure 18 -1A to 18 -3): Purified sequence optimized C50X-S53X variants of VHH 28D10 blocking hAng2-hTie2 (20-1), mAng2-mTie2 (20-2) and cAng2-cTie2 (20-3) interaction (ELISA).

Figure 19: Purified sequence optimized C50X-S53X variant of VHH 28D10 blocking hAngl-hTie2 interaction (ELISA).

Figure 20 (Figure 20-1A to 20-3C): Purified cycle 2 sequence optimized variants of VHH 28D10 blocking hAng2-hTie2 (22-1), mAng2-mTie2 (22-2) and cAng2-cTie2 (22-3) interaction (ELISA).

Examples

Materials and methods a) Generation of HEK293H stable cell lines overexpressing human or mouse Tie2 receptor [0175] The cDNAs encoding human Tie2 (NM_000459.3; SEQ ID NO:182;), mouse Tie2 (NM_013690.2; SEQ ID NO:183) and cyno Tie2 (SEQ ID NO:184); are cloned in pcDNA3.1-neo expression vector (Invitrogen, Carlsbad, CA, USA). To establish Human Embryonic Kidney (HEK) cells overexpressing human Tie2 or mouse Tie2, parental HEK293H cells undergo lipid mediated transfection with Fugene (Roche) with pcDNA3.1-neo-hTie2 or pcDNA3.1-neo-mTie2, respectively. For all conditions, transfectants are selected 2 days post-transfection by adding 1 mg/ml_ geneticin (Invitrogen, Carlsbad, CA, USA). For human, mouse and cyno Tie2, final high expressing clones are selected by single cell sorting clones binding to PE labeled anti-human Tie2 (R&D Systems, Minneapolis, MN, US), PE labeled anti-mouse Tie2 (eBioscience, San Diego, CA, USA) and a 2-step goat-anti-human Tie2 (R&D Systems, Minneapolis, MN, US) followed by PE labeled donkey-anti-goat (Jackson ImmunoResearch, West Grove, PA, USA), respectively, using the FACSAria Cell Sorter (BD Biosciences, San Jose, CA, USA). b) Generation of HEK293T cell lines overexpressing mouse or cynomolgus Ang2 and production of recombinant mouse and cynomolgus Ang2 conditioned medium [0176] The cDNAs encoding N-terminally FLAG-tagged mouse Ang2 (NM_007426.3; SEQ ID SEQ ID NO: 185) and cynomolgus Ang2 (AB172643.1; SEQ ID NO: 186) are cloned in a pSecTag2B expression vector (Invitrogen, Carlsbad, CA, USA). Human Embryonic Kidney (HEK) cells transiently overexpressing mouse Ang2 or cynomolgus Ang2 are generated by lipid-mediated transfection (Fugene; Roche) of pSecTag2B-mAng2 or pSecTag2B-cAng2, respectively, in the HEK293T parental cell line. Productions are performed in 1.5 liter CF10 Bag, and 1.5L conditioned medium (CM) is collected 5 days post-transfection. c) Production of recombinant cynomolgus Tie2/Fc chimera in HEK293-F Cells [0177] The cDNA encoding for the extracellular domain of Tie-2 is subcloned into the expression plasmid pSecTag2b using appropriate restriction sites to generate an Fc-fusion protein. Transfection into HEK293-F cells (Invitrogen) is performed as described by the manufacturer using Megaprep (Qiagen) preparations of plasmids, Optimem-Medium (Invitrogen), 293-fectin (Invitrogen) at an initial cell density of 1 x 106 viable cells/mL with 1 pg plasmid DNA/106 cells. Transfected cells are cultivated in shaker flasks for 7 days at 37 °C. Conditioned Medium (CM) is harvested by centrifugation at 4000g for 10 min and filtered through a sterile filter (0.45pm membrane).

[0178] Fc-fusion proteins are purified using affinity chromatography by loading the CM at 5 ml/min onto a 5 ml MabSelect SuRe Protein A column equilibrated with DPBS. After a washing step with DPBS, bound Fc-protein is eluted with 10 mM sodium citrate buffer pH 3.0 and subsequently neutralized to pH 7.0 by adding 1M Tris/HCI pH 8.0. The purified protein is concentrated and buffer exchanged to DPBS with a Millipore Amicon Ultra (10 kDa molecular weight cuttoff) centrifugal concentrator. Presence of the protein is confirmed with standard analytical methods (electrophoresis with Experion Pro 260 kit- BioRad; mass spectrometry). The protein is further analysed by size-exclusion chromatography and the endotoxin-content is determined (Endosafe PTS kit - Charles River). d) Production of recombinant human, cynomolgus, mouse and rat Ang2-FLD in HEK293-F Cells [0179] Molecular cloning and cell culture is performed as described for Tie2-Fc-fusion protein.

For purification of His-tagged proteins the CM is loaded at 5 ml/min on a 2 ml Ni2+ chelating sepharose fast flow column (His-Trap - GE Healthcare Life Sciences) equilibrated with DPBS. After loading in the presence of 4% elution buffer (DPBS + 0.5% imidazol) to prevent unspecific binding, the column is washed with DPBS. Ang2-FLD-proteins are eluted from the column with DPBS containing 0.5% imidazol. Subsequently an ultrafiltration step was done for concentration and buffer exchange (10 kDa molecular withgt cut off). An aliquot of the protein is retained for analytical characterization as described for Tie2-Fc.

Example 1

Immunization with recombinant human Ang2 induces a humoral immune response in llama 1.1. Immunizations [0180] After approval of the Ethical Committee of the faculty of Veterinary Medicine (University Ghent, Belgium), 4 llamas (designated No. 406, 408, 454, 455) are immunized with 4 intramuscular injections (day 0: 50 pg, day 14: 20 pg, day 28: 17.5 pg and day 42: 17.5 pg dose) of recombinant human Ang2 (R&D Systems, Minneapolis, MN, US). The antigen is formulated in Complete Freund's Adjuvant for the prime injection at day 0 (Difco, Detroit, Ml, USA) and in Incomplete Freund's Adjuvant for the booster injections (Difco, Detroit, Ml, USA). 1.2. Evaluation of induced immune responses in llama [0181] To evaluate the induction of an immune response in the animals against human Ang2 by ELISA, sera are collected at day 0 (pre-immune), day 35 and day 46 (time of peripheral blood lymphocyte [PBL] collection). In short, 1 pg/mL of recombinant human Ang2 (R&D Systems, Minneapolis, MN, USA) is immobilized overnight at 4°C in a 96-well MaxiSorp plate (Nunc, Wiesbaden, Germany). Wells are blocked with a casein solution (PBS + 1% casein). After addition of serial serum dilutions, specifically bound immunoglobulins are detected using a horseradish peroxidase (HRP)-conjugated goat anti-llama immunoglobulin (Bethyl Laboratories Inc., Montgomery, TX, USA) and a subsequent enzymatic reaction in the presence of the substrate TMB (3,3',5,5'-tetramentylbenzidine) (Pierce, Rockford, IL, USA), showing that a significant antibody dependent immune response against human Ang2 is induced. The antibody response is mounted both by conventional and heavy-chain only antibody expressing B-cell repertoires, since bound immunoglobulins can be detected with antibodies specifically recognizing the conventional llama IgG 1 antibodies or the heavy chain only llama lgG2 or lgG3 antibodies. In all llamas injected with human Ang2, an antibody response is mounted by conventional and heavy chain only antibody expressing B-cells specifically against human Ang2. The Ang2 serum titer responses for each llama are depicted in Table 1.

Table 1: Antibody mediated specific serum response against recombinant human Ang2.

; Legend: (*) i Low (or +/-): 1,000 > HSDs/N>2 < 1,500 | i Moderate (or +): 1,500 > HSDs/N>2 < 13,500 i Good (or ++): 13,500 > HSDs/N>2 < 365,000 i Excellent (or +++): HSDs/N>2 > 365,000 | i (*) HSD, Highest Serum Dilution; S/N > 2, signal-to-noise ratio > 2 |

Example 2

Cloning of the heavy-chain only antibody fragment repertoires and preparation of phage [0182] Following the final immunogen injection, immune tissues as the source of B-cells that produce the heavy-chain only antibodies are collected from the immunized llamas. Typically, two 150 mL blood samples collected 4 and 10 days after the last antigen injection, and one lymph node biopsy, collected 4 days after the last antigen injection are collected per animal. From the blood samples, peripheral blood mononuclear cells (PBMCs) are prepared using Ficoll-Hypaque according to the manufacturer's instructions (Amersham Biosciences, Piscataway, NJ, USA). From the PBMCs and the lymph node biopsy (not prelevated from animal No. 406), total RNA is extracted, which is used as starting material for RT-PCR to amplify the VHH encoding DNA segments, as described in Example 3 (page 46) of WO 05/044858. For each immunized llama, a library is constructed by pooling the total RNA isolated from all collected immune tissues of that animal. In short, the PCR-amplified VHH repertoire is cloned via specific restriction sites into a vector designed to facilitate phage display of the VHH library. The vector is derived from pUC119 and contains the LacZ promoter, a M13 phage gill

protein coding sequence, a resistance gene for ampicillin or carbenicillin, a multiple cloning site and a hybrid glll-pelB leader sequence. In frame with the VHH coding sequence, the vector encodes a C-terminal c-myc tag and a His6 tag. Phage are prepared according to standard protocols and stored after filter sterilization at 4°C for further use.

Example 3

Selection of Ang2 specific VHHs via phage display [0183] VHH repertoires obtained from all llamas and cloned as phage library are used in different selection strategies, applying a multiplicity of selection conditions. Variables include i) the Ang2 protein format: biotinylated C-terminally His-tagged full length recombinant human Ang2 (R&amp;D Systems, Minneapolis, MN, USA) and C-terminally His-tagged full length mouse Ang2 (produced at GeneArt, now Invitrogen, Carlsbad, CA, USA), ii) the Ang2 presentation method: plates directly coated with mouse Ang2 or incubation in solution with biotinylated human Ang2 followed by capturing on neutravidin-coated plates, and iii) the antigen concentration. All selections are done in 96 well MaxiSorp plates (Nunc, Wiesbaden, Germany).

[0184] Multi-round selections are performed as follows: Ang2 preparations for solid and solution phase selection formats are presented as described above at multiple concentrations (biotinylated human Ang2: 50, 5, 0.5, 0.05 and 0.005 nM; mouse Ang2: 10, 1, 0.1 and 0.01 pg/mL). After 2h incubation with the phage libraries, followed by extensive washing, bound phages are eluted with trypsin (1 mg/ml_) for 15-30 minutes at room temperature. Trypsin activity is then immediately neutralized by applying 0.8 mM protease inhibitor ABSF. As background control, selections w/o antigen are performed in parallel. Phage outputs that show enrichment over background are used to infect E. coli. Infected E. coli cells are either used to prepare phage for the next selection round (phage rescue) or plated on LB agar plates (ampicillin + glucose2%) for analysis of individual VHH clones. In order to screen a selection output for specific binders or blockers, single colonies are picked from the agar plates and grown in 1 mL 96-deep-well plates. LacZ-controlled VHH expression is induced by adding IPTG (0.1-1mM final) in the absence of glucose. Periplasmic extracts (in a volume of -80 uL) are prepared according to standard protocols (as disclosed in for example WO 2006/040153 cited herein). Briefly, cultures were centrifuged for 15 minutes at 4,500 rpm. The pellet was frozen overnight or for 1 hour at -20°C. Next, the pellet was thawed at room temperature for 40 minutes, re-suspended in 15 ml peri buffer (50 mM NaHPO4, 300mM NaCI) and shaken for 1 hour. Periplasmic fraction was isolated by centrifugation for 20 minutes at 14000 rpm.

Example 4

Screening of periplasmic extracts in Ang2-Tie2 and Ang1-Tie2 ELISA and AlphaScreen competition assays [0185] Periplasmic extracts containing expressed VHHs are screened in a human Ang2-human Tie2 AlphaScreen competition assay to assess their blocking capacity. In brief, human Tie2/Fc chimera (R&amp;D Systems, Minneapolis, MN, USA) is biotinylated using /V-hydroxysulfosuccinimide ester of biotin (Thermo Fisher Scientific, Rockford, IL, USA). FLAG tagged human Ang2 (Alexis Biochemicals, San Diego, CA, USA) is captured using Acceptor beads (Perkin Elmer, Waltham, MA, US) coated with anti-FLAG M2 antibody (Sigma, St Louis, MO, USA). To evaluate the capacity of the VHHs to inhibit binding of human Ang2 to its receptor human Tie2, 1:25 dilutions of the periplasmic extracts containing expressed VHHs are incubated with 0.1 nM FLAG tagged human Ang2. To this mixture, the Acceptor beads and 0.3 nM biotinylated human Tie2/Fc chimeras are added and further incubated for 2 hours at room temperature. Finally, streptavidin conjugated Donor beads (Perkin Elmer, Waltham, MA, US) are added and the mixture is incubated for an additional 2 hours at room temperature. Assay buffer is PBS + 0.03% Tween-20 + 0.1% BSA. Fluorescence is measured using the Envision Multilabel Plate reader (Perkin Elmer, Waltham, MA, USA) using an excitation wavelength of 680 nm and an emission wavelength of 520 nm. Decrease in fluorescence signal indicates that the binding of human Ang2 to human Tie2 is blocked by the VHH expressed in the periplasmic extract. VHHs able to block the human Ang2-human Tie2 interaction for at least 50% are screened in a confirmatory ELISA based competition assay. Additionally, cross-reactivity for binding to mouse Ang2 and selectivity over human Ang1 is also assessed in a competition ELISA. In brief, human or mouse Tie2/Fc chimera (R&amp;D Systems, Minneapolis, MN, USA) are immobilized at 2 pg/mL overnight at 4°C in a 96-well MaxiSorp plate (Nunc, Wiesbaden, Germany). Wells are blocked with a 1% casein solution. A 1:5 dilution of periplasmic extract containing expressed VHHs is incubated with the following Ang species according to the type of assay: 0.02 nM FLAG tagged human Ang2, a 1:3,000 dilution of HEK293 conditioned medium containing FLAG tagged mouse Ang2 or 0.02 nM FLAG-tagged human Ang1 (Alexis Biochemicals, San Diego, CA, USA). This mixture is added to the Tie2/Fc coated well and incubated for 2 hours at room temperature.

[0186] Residual binding of Ang is detected using HRP-conjugated anti-FLAG M2 antibody (Sigma, St Louis, MO, USA).

[0187] In a second screening cycle, periplasmic extracts containing expressed VHHs of selection outputs that yielded a high diversity of mouse Ang2 cross-reactive blocking VHHs are screened at a 1:300 dilution. VHHs inhibiting the binding of human Ang2 to human Tie2, mouse Ang2 to mouse Tie2 and showing no inhibition of human Ang1 binding to human Tie2 are selected. Sequence analysis revealed 86 unique VHHs belonging to 38 different B-cell lineages. The total number of unique sequence variants found for each B-cell lineage, a representative VHH and the selection condition used, is depicted in Table 2. An overview of AlphaScreen and ELISA based screening data is given in Table 3. The amino acid sequences of all unique VHHs are shown in the Sequence Listing (SEQ ID NOs: 1 to 86) and in Table 4.

Table 2: Selection parameters used for the identification of Ang2 specific VHH B-cell lineages.

Table 3: Screening of periplasmic extracts containing expressed anti-Ang2 VHH(a)

; (a)if multiple unique VHH variants within a B-cell lineage are identified, the range (min-max) of % inhibition is given.

Table 4: Amino acid sequence of unique anti-Ang2 VHHs identified during screening

Example 5

Characterization of purified anti-Ang2 VHHs [0188] A subset of inhibitory anti-Ang2 VHHs selected from the screening described in Example 4 are further purified and characterized. Selected VHHs are expressed in E. coli TG1 as c-myc, His6-tagged proteins. Expression is induced by addition of 1 mM IPTG and allowed to continue for 4 hours at 37°C. After spinning the cell cultures, periplasmic extracts are prepared by freeze-thawing the pellets. These extracts are used as starting material and VHHs are purified via IMAC and size exclusion chromatography (SEC) resulting in > 95% purity as assessed via SDS-PAGE.

5.1. Evaluation of hAng2 blocking VHHs in ELISA

[0189] The blocking capacity of the VHHs is evaluated in a human Ang2-human Tie2 blocking ELISA. In brief, 2 pg/mL of Tie2/Fc chimera (R&amp;D Systems, Minneapolis, MN, USA) is coated in a 96-well MaxiSorp plate (Nunc, Wiesbaden, Germany). A fixed concentration of 0.02 nM FLAG-tagged human Ang2 (Alexis Biochemicals, San Diego, CA, USA) is added to a dilution series of the purified VHH (diluted in PBS+0.1% casein+0.05% Tween-20), and incubated on the coated human Tie2 receptor for 2 hours. Residual binding of human Ang2 is detected using horseradish peroxidase (HRP) conjugated anti-FLAG M2 (Sigma, St. Louis, MO, USA) (Figure 1). Reference molecule is the Fab fragment of Ab536 (US2009/0191212) (Figure 1-1) or the peptide moiety of peptibody AMG386 (SEQ ID NO:25 in W02004/092215) (Figure 1-2). As negative control an irrelevant VHH is used. The IC50 values for VHHs blocking the human Ang2-human Tie2 interaction are depicted in Table 5-1 and Table 5-2, respectively.

Table 5-1: IC50 (nM) values of purified VHHs blocking the hAng2/hTie2 interaction (competition ELISA; VHH: n=2-3; Fab Ab536: n=6)

Table 5-2: IC50 (nM) values of purified VHHs blocking the hAng2-hTie2 interaction (competition ELISA; VHH: n=1-3; AMG386 peptide: n=3)

5.2. Evaluation of cross-reactivity towards mouse and cynomolgus Ang2 in blocking ELISA

[0190] In order to determine if the VHH inhibits binding of mouse Ang2 to mouse Tie2 and cyno Ang2 to cyno Tie2, a competition ELISA is performed. In brief, 2 pg/mL of recombinant mouse Tie2-Fc or cyno Tie2-Fc is coated overnight at 4°C in a 96-well MaxiSorp plate (Nunc, Wiesbaden, Germany). Coated wells are blocked with a 1% casein solution. FLAG-tagged mouse Ang2 (1:3,000 dilution of conditioned medium from transient HEK transfection) or FLAG-tagged cyno Ang2 (1:800 dilution of conditioned medium from transient HEK transfection) and a dilution series of purified VHH (diluted in PBS+0.1% casein+0.05% Tween-20) are incubated on the coated Tie2-Fc receptor for 2 hours at room temperature to reach binding equilibrium. Residual binding of FLAG-mAng2 or FLAG-cAng2 is detected using HRP conjugated anti-FLAG

M2 mAb (Sigma, St. Louis, MO, USA). Reference molecule is the Fab fragment of Ab536 (mouse: Figure 2-1) or the peptide moiety of peptibody AMG386 (mouse: Figure 2-2; cyno: Figure 3). As negative control an irrelevant VHH is used. The IC50 values for VHHs blocking the mouse Ang2-mouse Tie2 interaction are depicted in Table 6-1. The IC50 values for VHHs blocking the mouse and cyno Ang2 binding to mouse and cyno Tie2, respectively, is shown in Table 6-2.

Table 6-1: IC50 (nM) values of purified VHHs blocking the interaction of mAng2 to mTie2 (competition ELISA; VHH: n=2-3; Fab Ab536: n=5)

Table 6-2: IC50 (nM) values of purified VHHs blocking the interaction of mAng2 and cAng2 to mTie2 and cTie2, respectively (competition ELISA; VHH: n=1-3; AMG386 peptide: n=3; n.d., not determined)

5.3. Evaluation of selectivity of human Ang2 blocking VHHs towards human Ang1 in ELISA

[0191] In order to determine whether the anti-Ang2 blocking VHHs are selective over human Ang1 binding to human Tie2, a competition ELISA is performed. In brief, 2 pg/mL of recombinant human Tie2-Fc (R&amp;D Systems, Minneapolis, MN, USA) is coated overnight at 4°C in a 96-well MaxiSorp plate (Nunc, Wiesbaden, Germany). Coated wells are blocked with a 1% casein solution. A fixed concentration (0.02 nM) of FLAG-tagged recombinant human Ang1 (Alexis Biochemicals, San Diego, CA, USA) and a dilution series of VHH (diluted in PBS+0.1 %casein+0.05%Tween-20) are incubated on the coated receptor Tie2-Fc for 2 hours at room temperature to reach binding equilibrium. Residual binding of FLAG-hAng1 is detected using HRP conjugated anti-FLAG M2 mAb. Reference molecule is the peptide moiety of AMG386 peptibody (Figure 4). As negative control an irrelevant VHH is used. The indicative IC50 values for VHHs blocking the human Ang1 - human Tie2 interaction are depicted in Table 7.

Table 7: IC50 (nM) values of purified VHHs blocking the interaction of human Ang1 to human Tie2 (competition ELISA; VHH: n=2-3; AMG386 peptide: n=3)

5.4. Determining the affinity of the human, mouse, cyno Ang2 - VHH interaction [0192] Affinities of the VHH for binding to human, mouse and cyno Ang2 are determined using surface plasmon resonance (SPR) analysis (Biacore T100). In brief, VHH and benchmark compounds are immobilized on a CM5 chip via amine coupling. A multi-cycle kinetic approach is used: different concentrations of human, mouse and cyno Ang2-FLD (0.4-1-2.6-6.4-16-40-100 nM) are injected. Ang2-FLD species are allowed to associate for 2 min and to dissociate for 20 min at a flow rate of 45 pL/min. In between injections, the surfaces are regenerated with a 10 sec pulse of 25 mM NaOH and 60 sec stabilization period. Association/dissociation data are evaluated by fitting a 1:1 interaction model (Langmuir binding). The affinity constant «□ is calculated from resulting association and dissociation rate constants ka and kj and are depicted in Table 8.

Table 8: Affinity Kq (nM) of purified VHHs for human, mouse and cyno Ang2

Example 6

Affinity maturation of selected VHH

[0193] A variant of VHH 28D10 (00027 carrying C50S/S53N and Q-iosL substitution - Example 7.3) is subjected to affinity maturation.

[0194] In a first cycle, amino acid substitutions are introduced randomly in both framework (FW) and complementary determining regions (CDR) using the error-prone PCR method. Mutagenesis is performed in a two-round PCR-based approach using the Genemorph II Random Mutagenesis kit (Stratagene, La Jolla, CA, USA) using 1 ng of VHH 00027 cDNA template, followed by a second error-prone PCR using 0.1 ng of product of round 1. After a polish step, PCR products are inserted via unique restriction sites into a vector designed to facilitate phage display of the VHH library. Consecutive rounds of in-solution selections are performed using decreasing concentrations of biotinylated recombinant human Ang2 (R&amp;D Systems, Minneapolis, MN, USA) and trypsin elutions. Periplasmic extracts (in a volume of ~ 80 uL) are prepared according to standard methods and screened for binding to recombinant human Ang2-FLD in a ProteOn (BioRad, Hercules, CA, USA) off-rate assay. In brief, a GLC ProteOn Sensor chip is coated with recombinant human Ang2-FLD on the "ligand channels" L3, L4, L5 and L6 (with L1/L2 as reference channel). Periplasmic extract of affinity matured clones is diluted 1:10 and injected across the "analyte channels" A1-A6. An average off-rate is calculated of the reference VHH 00027 which is prepared and tested in the same way as the affinity matured VHHs and serves as a reference to calculate off-rate improvements. The top 25 affinity matured variants are shown in Table 9. VHH are sequenced (Table 10-A) to identify amino acid mutations beneficial for improving the off-rate (Table 10-B).

Table 9: Off-rate and fold improvement of affinity matured variants of VHH 00027.

[0195] Initially 12 VHH variants containing combinations of mutations on Kabat position 27, 29, 100b and 100i (Table 11; Figure 5) are constructed. The different combinations of these 4 mutations are grafted on the sequence optimized VHH 00042 backbone (Figure 17-B) containing an additional D54G substitution (Example 6.3). The amino acid sequence is aligned to the human germline VH3/JH consensus sequence. Residues are numbered according to Kabat, CDRs are shown in grey according to AbM definition (Oxford Molecular's AbM antibody modeling software). Constructs are cloned into the expression vector pAX100 in frame with a C-terminal c-myc tag and a (His)6 tag. VHH variants are produced in E. coli and purified by IMAC and SEC. Sequences are represented in Table 11. All these VHH are analysed in the hAng2/hTie2 (Example 5.1; results shown in Figure 6 and Table 12), and hAng1/hTie2 competition ELISA (Example 5.3; results shown in Figure 7 and Table 12). Additionally, the melting temperature (Tm) of each variant at pH7 is determined in a thermal shift assay, which is based on the temperature dependent change in fluorescence signal upon incorporation of Sypro Orange (Invitrogen, Carlsbad, CA, USA) (Ericsson et al, Anal. Biochem. 357 (2006), pp289-298) (Table 12).

Table 10-A: Amino acid sequence of affinity matured anti-Ang2 VHHs

Table 10-B: Single mutations or combination thereof yielding improvements in off-rate

Table 11: Amino acid sequence of affinity matured anti-Ang2 VHHs

Table 12: Overview of Tm, IC50 (pM) in human, mouse and cyno Ang2 competition ELISA and hAng1/hAng2 IC50 ratios of affinity matured variants of VHH 00027

[0196] From potency, Tm and sequence perspective VHH 00908 is taken forward into a second cycle of combined affinity maturation and sequence optimization (Example 7.3).

Example 7

Sequence optimization of selected VHH 1D01, 28D10 and 37F02 7.1 VHH 1D01 [0197] The amino acid sequence of anti-Ang2 VHH 1D01 (see Figure 8-A) is aligned to the human germline VH3/JH consensus sequence. Residues are numbered according to Kabat, CDRs are shown in grey according to AbM definition (Oxford Molecular's AbM antibody modeling software). Residues to be mutated to their human counterpart are underlined. Potential post-translational modification sites to be tackled are boxed. The alignment shows that 1D01 contains 6 framework mutations relative to the reference germline sequence. Non-human residues at positions 14, 41, 71, 74, 83 and 108 are selected for substitution with their human germline counterpart. Aset of seven 1D01 variants carrying different combinations of human residues on these positions (Figure 8-B) is constructed and produced (Example 5). In parallel, in 3 of these 7 variants a potential Asp isomerization site at position D54G55 is removed by

introducing a D54G substitution, and in 1 of these 7 variants a potential pyroGlu formation site at position E-| is removed by an E-|D substitution (AA sequences are listed in Table 15).

[0198] These variants are characterized as purified protein in the human (Figure 9-1), mouse (Figure 9-2) and cyno (Figure 9-3) Ang2/Tie2 competition ELISA (Example 5.1; Example 5.2), the hAng1/hTie2 competition ELISA (Example 5.3; Figure 10). Additionally, melting temperature (Tm) of each variant is determined in thermal shift assay (Example 6). An overview of the data can be found in Table 13. Additionally, % FR identity to the human germline is calculated according to AbM definition (Oxford Molecular's AbM antibody modeling software). Affinity of VHH 00921 for human, cyno, mouse and rat Ang2 is shown in Table 14 (Example 5.4).

Table 13: Overview of Tm, IC50 (nM) in human, mouse and cyno Ang2 competition ELISA and hAng1/hAng2 IC50 ratios of sequence optimized variants of VHH 1D01

i n.d., not determined |

Table 14: Affinity «□ of purified VHH 00921 for recombinant human, cyno, mouse and rat Ang2

Table 15: Amino acid sequence of sequence optimized variants of anti-Ang2 VHH 1D01

7.2 VHH 37F02 [0199] The amino acid sequence of anti-Ang2 VHH 37F02 (see Figure 12-A) is aligned to the human germline VH3/JH consensus sequence. Residues are numbered according to Kabat, CDRs are shown in grey according to AbM definition (Oxford Molecular's AbM antibody modeling software). Residues to be mutated to their human counterpart are underlined. Potential post-translational modification sites to be tackled are boxed. The alignment shows that 37F02 contains 4 framework mutations relative to the reference germline sequence. Nonhuman residues at positions 60, 74, 83 and 108 are selected for substitution with their human germline counterpart. In parallel, a potential Asp isomerization site at position D54G55 is removed by introducing a D54G substitution. A set of three cycle 1 37F02 variants carrying different combinations of human residues on these positions (Figure 12-B) is constructed and produced (Example 5; AA sequences are listed in Table 21-1).

[0200] These variants are characterized as purified protein in the human (Figure 12-1), mouse (Figure 12-2) and cyno (Figure 12-3) Ang2/Tie2 competition ELISA (Example 5.1; Example 5.2). Additionally, melting temperature (Tm) of each variant is determined in thermal shift assay (Example 6). An overview of the data can be found in Table 16. Additionally, % FR identity to the human germline is calculated according to AbM definition (Oxford Molecular's AbM antibody modeling software).

Table 16: Overview of Tm, IC50 (pM) in human, mouse and cyno Ang2 competition ELISA and hAng1/hAng2 IC50 ratios of cycle 1 sequence optimized variants of VHH 37F02

[0201] A NNK library approach is used to knock out two potential post-translational

modifications sites in CDR3: i) oxidation sensitive Met on position 100e and ii) Asp isomerization site on position D95S96. Since D54G is tolerated (VHH 00046 and 00920; Table 16 and Table 17) no NNK approach was used to knock out this potential Asp isomerization site.

[0202] In the end, 3 NNK libraries containing VHH clones carrying substitutions at positions D95, S96 and M-|ooe to θϋ other amino acids are screened in a hAng2/hTie2 competition AlphaScreen assay (Example 2). Briefly, periplasmic extracts containing expressed VHH are screened at 3 different dilutions (corresponding roughly to EC20, EC50 and ΕΟθο of the parental VHH 37F02) and changes in % inhibition at the different dilution points are compared to parental 37F02. Based on the screening results and the data shown in Table 17, 8 additional cycle 2 VHH variants are constructed (based on VHH 00920 backbone) carrying different knock-out combinations of D95S96 and M-iooe (Figure 11-C; AA sequences are listed in Table 19-2).

[0203] All these variants are characterized as purified protein in the human (Figure 13-1), mouse (Figure 12-2) and cyno (Figure 12-3) Ang2/Tie2 competition ELISA (Example 5.1; Example 5.2), the hAng1/hTie2 competition ELISA (Example 5.3; Figure 14). Additionally, melting temperature (Tm) of each variant is determined in thermal shift assay (Example 6). An overview of the data can be found in Table 17. Additionally, % FR identity to the human germline is calculated. Affinity of VHH 00928 for human, mouse, cyno and rat Ang2 is shown in Table 18.

Table 17: Overview of Tm, IC50 (pM) in human, mouse and cyno Ang2 competition ELISA and hAng1/hAng2 IC50 ratios of cycle 2 sequence optimized variants of VHH 37F02

i n.d., not determined |

Table 18: Affinity Kq of purified VHH 00928 for recombinant human, cyno, mouse and rat Ang2

7.3 VHH 28010 [0204] The amino acid sequence of anti-Ang2 VHH 28D10 (see Figure 15-A) is aligned to the human germline VH3/JH consensus sequence. Residues are numbered according to Kabat, CDRs are shown in grey according to AbM definition (Oxford Molecular's AbM antibody modeling software). Residues to be mutated to their human counterpart are underlined. Potential post-translational modification sites to be tackled are boxed. The alignment shows that 28D10 contains 5 framework mutations relative to the reference germline sequence. Nonhuman residues at positions 14, 71, 74, 83 and 108 are selected for substitution with their human germline counterpart. In parallel, a potential Asp isomerization site at position D54G55 is removed by introducing a D54G substitution. The free cystein at position 50 was removed by substitution with Ala, Thr or Ser. In the end, a set of eleven cycle 1 28D10 variants carrying different combinations of human residues on these positions is constructed and produced (see Figure 15-B; AA sequences are listed in Table 24-1).

[0205] These variants are characterized as purified protein in the human (Figure 16-1), mouse (Figure 16-2) and cyno (Figure 16-3) Ang2/Tie2 competition ELISA (Example 5.1; Example 5.2), the hAng1/hTie2 competition ELISA (Example 5.3; Figure 17). Additionally, melting temperature (Tm) of each variant is determined in thermal shift assay (Example 6). An overview of the data can be found in Table 20. Additionally, % FR identity to the human germline is calculated.

Table 20: Overview of Tm, IC50 (nM) in human, mouse and cyno Ang2 competition ELISA and hAng1/hAng2 IC50 ratios of cycle 1 sequence optimized variants of VHH 28D10

n.d., not determined | [0206] An additional set of variants (cycle 2) is created to explore the affinity maturation substitution on position A24V and to further explore C50X-S53X variants (Table 21; Figure 15-C; AA sequences are listed in Table 23-4-2). These variants are characterized as purified protein in the human (Figure 18-1), mouse (Figure 18-2) and cyno (Figure 18-3) Ang2/Tie2 competition ELISA (Example 5.1; Example 5.2), the hAng1/hTie2 competition ELISA (Example 5.3; Figure 19). Additionally, melting temperature (Tm) of each variant is determined in thermal shift assay (Example 6).

Table 21: Overview of Tm, IC50 (nM) in human, mouse and cyno Ang2 competition ELISA and hAng1/hAng2 IC50 ratios of cycle 2 sequence optimized variants of VHH 28D10

ί n.d., not determined | [0207] In parallel, a NNK library approach is used to knock out two post-translational modifications sites in CDR2: two sequential Asp isomerization sites on position D52aS53 and D54G55. Since D54G is tolerated (Example 6; Table 20) no NNK approach was used to knock out this potential Asp isomerization site. In the end, 2 NNK libraries containing VHH clones carrying substitutions at positions D52a and S53 to all other amino acids are screened in a hAng2/hTie2 competition AlphaScreen assay (Example 2). Briefly, periplasmic extracts containing expressed

VHH are screened at 3 different dilutions (corresponding roughly to EC20, EC50 and ΕΟθο of the reference 00902) and changes in % inhibition at the different dilution points are compared to reference 00902. Based on the screening results and the data shown in Table 21, 7 additional VHH cycle 3 variants are constructed (based on 00908 backbone) (see Figure 15-D). The final aim is to construct VHH variants that retain or show increased potency, increased thermostability and have relevant PTM sites knocked out compared to VHH 28D10. (AA sequences are listed in Table 24-3). These variants are characterized as purified protein in the human (Figure 20-1), mouse (Figure 20-2) and cyno (Figure 20-3) Ang2/Tie2 competition ELISA (Example 5.1; Example 5.2), the hAng1/hTie2 competition ELISA (Example 5.3). Additionally, melting temperature (Tm) of each variant is determined in thermal shift assay (Example 6). An overview of the data can be found in Table 22. Additionally, % FR identity to the human germline is calculated. The most optimal sequence changes were finally applied to a non-affinity matured variant VHH 00956 (Figure 15-D). Affinity of VHH 00919, 00938 and 00956 for human, mouse, cyno and rat Ang2 is shown in Table 23.

Table 22: Overview of Tm, IC50 (nM) in human, mouse and cyno Ang2 competition ELISA and hAng1/hAng2 IC50 ratios of cycle 3 sequence optimized variants of VHH 28D10

ί n.d. not determined |

Table 23: Affinity Kq of purified VHHs 00919, 00938 and 00956 for recombinant human, cyno, mouse and rat Ang2

Table 24-2

[0208] VHH domain 00938 is an embodiment of the invention as claimed in the appended claims. Other VHH domains, shown above but not falling under these claims, are shown for comparative purposes or to explain how VHH domain 00938 was obtained.

SEQUENCE LISTING

[0209]

<110> Boehringer Ingelheim International GmbH

<120> ANG2-BINDING MOLECULES

<130> 12-0350-PCT <160> 190 <170> Patentln version 3.3 <210> 1 <211> 124

<212> PRT <213> Lama glama <400> 1

Glu Val Gin Leu Val Glu Ser Gly Gly Gly Leu Val Gin Ala Gly Gly 15 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Asp Asp Tyr 20 25 30

Ala Leu Gly Trp Phe Arg Gin Ala Ala Gly Lys Glu Arg Glu Gly Val 35 40 45

Ser Cys He Arg Cys Ser Asp Gly Ser Thr Tyr Tyr Ala Asp Ser Val 50 55 60

Lys Gly Arg Phe Thr lie Ser Ser Asp Asn Ala Lys Asn Thr Val Tyr 65 70 75 80

Leu Gin Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Ala Ser He Val Pro Arg Ser Lys Leu Glu Pro Tyr Glu Tyr Asp 100 105 110

Ala Trp Gly Gin Gly Thr Gin Val Thr Val Ser Ser 115 120 <210> 2 <211> 111

<212> PRT <213> Lama glama <400>2

Glu Val Gin Leu Val Glu Ser Gly Gly Gly Leu Val Gin Pro Gly Gly 15 10 15

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Ala Thr Phe Thr Asn Arg Gly Ser Thr Tyr Tyr Ala Gly Ser Val Lys 50 55 60

Gly Arg Phe Thr He Ser Arg Asp Asn Ala Lys Asn Thr Met Tyr Leu 65 70 75 80

Gin Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Val Tyr Tyr Cys Asn 85 90 95

Thr Gly Pro Tyr Trp Gly Gin Gly Thr Gin Val Thr Val Ser Ser 100 105 HO <210> 3 <211> 120

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Glu val Gin Leu Val Glu Ser Gly Gly Gly Leu Val Gin Ala Gly Gly 15 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Arg Thr Phe Ser Ser Ser 20 25 30

Val Met Gly Trp Phe Arg Gin Ala Pro Gly Lys Glu Arg Glu Phe Val 35 40 45

Ala Ala lie Ser Gly Ser Gly Ser Ser Thr Asp Ser Ala Gin Gly Arg 50 55 60

Phe Thr lie Ser Arg Asp Asn Ala Lys Asn Thr Val Tyr Leu Gin Met 65 70 75 80

Asn Ser Leu Lys Pro Glu Asp Thr Ala Val Tyr Tyr Cys Ala Ala Gly 85 90 95

Arg Ala Phe Leu Ala Arg Asp Thr Phe Tyr Tyr Asp lie Trp Gly Gin 100 105 110

Gly Thr Gin Val Thr Val Ser Ser 115 120 <210> 4 <211> 126

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Glu Val Gin LeU Val Glu Ser Gly Gly Gly Leu Val Gin Ala Gly Gly 15 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Ala Phe Asp Asp His 20 25 30

Leu lie Gly Trp Phe Arg Gin Ala Pro Gly Lys Glu Arg Glu Ala Val 35 40 45

Ser Cys lie Ser Arg Ser Ala Gly Ser Thr Tyr Tyr Ala Asp Ser Val 50 55 60

Lys Gly Arg Phe Thr lie Ser Ser Asp Asn Ala Lys Asn Thr Val Tyr 65 70 75 80

Leu Gin Met Asn Ser Leu Lys Ala Asp Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Ala Gly Pro Ala Trp Gly Arg Pro Ala Ser Pro Leu Pro Tyr Glu 100 105 110

Tyr Asp Tyr Trp Gly Gin Gly Thr Gin Val Thr Val Ser Ser 115 120 125 <210> 5 <211> 134

<212> PRT <213> Lama glama <400>5

Glu Val Gin Leu Val Glu Ser Gly Gly Gly Leu Val Gin Ala Gly Gly 1 5 10 15

Ser Leu Arg Leu Ser Gys Ala Ala Ser Gly Arg Thr lie Ser Ser Tyr 20 25 30

Ala Met Ala Trp Phe Arg Gin Ala Pro Gly Lys Glu Arg Glu Phe Val 35 40 45

Ala Ala lie Ser Leu Ser Gly Asp Ser Thr Tyr Tyr Ala Asp Ser Val 50 55 60

Lys Gly Arg Phe Thr lie Ser Arg Asp Asn Ala Lys Asn Thr Val Tyr 65 70 75 80

Leu Gin Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Glu Tyr Tyr Cys 85 90 95

Ala Ala Thr Asp Trp Asp Phe Glu Asp lie Pro Glu Tyr Tyr Cys Ser 100 105 110

Gly Tyr Gly Cys Asp Glu Ser Leu Phe Asp Ser Trp Gly Gin Gly Thr 115 120 125

Gin Val Thr Val Ser Ser 130 <210> 6 <211> 130

<212> PRT <213> Lama glama <400> 6

Glu Val Gin Leu Val Glu Ser Gly Gly Gly Leu Val Gin Pro Gly Gly 15 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Leu Asp Tyr Asp 20 25 30

Ala lie Gly Trp Phe Arg Gin Ala Pro Gly Lys Glu Arg Glu Gly Val 35 40 45

Ser Cys lie Thr Ser Ser Asp Gly lie Thr Tyr Tyr Ala Asp Ser Val 50 55 60

Lys Gly Arg Phe Thr lie Ser Arg Asp Ash Ala Lys Asn Thr Val Tyr 65 70 75 80 Τλπ m τί Mdf lah Sar Τ.απ Τ,νβ Pra Glu larS Thr ft! a Val Twr Twr Pva 85 "" 90*” 95

Ala Thr Gly Asn Arg Arg lie Tyr Tyr Ser Asp Tyr Ala Leu Ala Cys 100 105 110

Phe Pro Tyr Glu Tyr Asp Tyr Trp Gly Gin Gly Thr Gin Val Thr Val 115 120 125

Ser Ser 130 <210> 7 <211> 123

<212> PRT <213> Lama glama <400>7

Glu Val Gin Leu Val Glu Ser Gly Gly Gly Leu Val Gin Val Gly Asp 1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Arg Thr Phe Ser Thr Tyr 20 25 30

Leu Met Val Gly Trp Phe Arg Gin Ala Pro Gly Lys Glu Arg Glu Phe 35 40 45

Ala Ala Val He Trp Ser Ser Gly Asp Thr Ala Tyr Ala Asp Ser Val 50 55 60

Lys Gly Arg Phe Thr He Ser Arg Asp Ser Ala Lys Asn Thr Val Tyr 65 70 75 80

Leu Gin Met Asn Ser Leu Lys Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Gly Ser Tyr Asp Gly Asn Tyr Tyr He Pro Gly Phe Tyr Lys Asp 100 105 110

Trp Gly Gin Gly Thr Gin Val Thr Val Ser Ser 115 120 <210> 8 <211> 130

<212> PRT <213> Lama glama <400> 8

Glu Val Gin Leu Val Glu Ser Gly Gly Gly Leu Val Gin Pro Gly Gly 15 10 15

Ser Leu Arg Leu Ser Cys Thr Thr Ser Gly Phe Thr Leu Asp Tyr Tyr 20 25 30

Ala Val Gly Trp Phe Arg Gin Ala Pro Gly Lys Glu Arg Glu Gly Val 35 40 45

Ser Cys lie Ser Ser Ser Asp Gly Ser Thr Tyr Tyr Ala Asp Ser Val 50 55 60

Lys Gly Arg Phe Thr lie Ser Arg Asp Asn Ala Lys Asn Thr Val Tyr 65 70 75 80

Leu Gin Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Thr Gly Asn Leu Arg He Tyr Tyr Ser Asp Tyr Ala Leu Ala Cys 100 105 110

Phe Pro Tyr Glu Tyr Asp Tyr Trp Gly Gin Gly Thr Gin Val Thr Val 115 120 125

Ser Ser 130 <210> 9 <211> 123

<212> PRT <213> Lama glama <400>9

Glu Val Gin Leu Val Glu Ser Gly Gly Gly Leu Val Gin Val Gly Asp 15 10 15

Ser Leu Arg Leu Ala Cys Ala Ala Ser Gly Arg Thr Phe Ser Thr Tyr 20 25 30

Leu Met Val Gly Trp Phe Arg Gin Ala Pro Gly Lys Glu Arg Glu Phe 35 40 45

Ala Ala Gly lie Trp Ser Ser Gly Asp Thr Ala Tyr Ala Asp Ser Val 50 55 60

Lys Gly Arg Phe Thr He Ser Arg Asp Asn Ala Lys Asn Thr Val Tyr 65 70 75 80

Leu Gin Met Asn Ser Leu Lys Thr Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Gly Ser Tyr Asp Gly Asn Tyr Tyr He Pro Gly Phe Tyr Lys Asp 100 105 110

Trp Gly Gin Gly Thr Gin Val Thr Val Ser Ser 115 120 <210> 10 <211> 123

<212> PRT <213> Lama glama <400> 10

Glu Val Gin Leu Val Glu Ser Gly Gly Gly Leu Val Gin Val Gly Asp 1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Arg Thr Phe Ser Thr Tyr 20 25 30

Leu Met Val Gly Trp Phe Arg Gin Ala Pro Gly Lys Glu Arg Glu Phe 35 40 45

Ala Ala Ala Met Trp Ser Ser Gly Val Pro Ala Tyr Ala Asp Ser Val 50 55 60

Lys Gly Arg Phe Thr He Ser Arg Asp Asn Ala Lys Asn Thr Val Tyr 65 70 75 80

Leu Gin Met Asn Ser Leu Lys Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Gly Ser Tyr Gly Gly Asn Tyr Tyr He Pro Gly Phe Tyr Glu Asp 100 105 110

Trp Gly Gin Gly Thr Gin Val Thr Val Ser Ser 115 120 <210> 11 <211> 124

<212> PRT <213> Lama glama <400> 11

Glu Val Gin LeU Val Glu Ser Gly Gly Gly Leu Val Gin Ala Gly Gly 15 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly His Thr Phe Ser Arg Tyr 20 25 30

Ala Met Gly Trp Phe Arg Arg Val Pro Gly Lys Glu Arg Glu Phe Val 35 40 45

Thr His He Thr Trp Asn Arg Gly Ser Thr Tyr Tyr Ala Asp Ser Val 50 55 60

Lys Gly Arg Phe Thr He Ser Arg ASp Lys Ala Ser Asn Thr Leu Tyr 65 70 75 80

Leu Gin Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Val Tyr Tyr CyS 85 90 95

Ala Ala Gin He Lys Tyr Gly Ala Val Thr His Pro Glu Glu Tyr Ser 100 105 110

Tyr Trp Gly Gin Gly Thr Gin Val Thr Val Ser Ser 115 120 <210> 12 <211> 126

<212> PRT <213> Lama glama <400> 12

Lys Val Gin Leu Val Glu Ser Gly Gly Gly Leu Val Gin Ala Gly Gly 15 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Ala Phe Asp Asp His 20 25 30

Leu lie Gly Trp Phe Arg Gin Ala Pro Gly Lys Glu Arg Glu Ala Val 35 40 45

Ser Cys lie Ser Arg Ser Ala Gly Ser Thr Tyr Tyr Ala Asp Ser Val 50 55 60

Lys Gly Arg Phe Thr lie Ser Ser Asp Asn Ala Lys Asn Thr Val Tyr 65 70 75 80

Leu Gin Met Asn Ser Leu Lys Ala Asp ASp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Ala Gly Pro Ala Trp Gly Arg Pro Ala Ser Pro Leu Pro Tyr Glu 100 105 110

Tyr Asp Tyr Trp Gly Gin Gly Thr Gin Val Thr Val Ser Ser 115 120 125 <210> 13 <211> 134

<212> PRT <213> Lama glama <400> 13

Glu Val Gin Leu Val Glu Ser Gly Gly Gly Leu Val Gin Ala Gly Gly 15 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Asp Asp Tyr 20 25 30

Ala lie Gly Trp Phe Arg Gin Ala Pro Gly Lys Glu Arg Glu Gly Val 35 40 45

Ser Cys He Ser Ser Ser Asp Gly Ser Thr Ala Tyr Ala Asp Ser Val 50 55 60

Lys Gly Arg Phe Thr lie Ser Ser Asp Asn Ala Lys Asn Thr Val Tyr 65 70 75 80

Leu Gin Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Thr Ala val Pro Ala Thr Arg Arg Thr Pro Gin Met Val Ala Ala Asn 100 105 110

Val Cys Trp Leu Ala Glu Tyr Glu Tyr Asp Tyr Trp Gly Gin Gly Thr 115 120 125

Gin Val Thr Val Ser Ser 130 <210> 14 <211> 123

<212> PRT <213> Lama glama <400> 14

Glu val Gin Leu Val Glu Ser Gly Gly Gly Leu Val Gin Val Gly Asp 15 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Arg Thr Phe Ser Thr Tyr 20 25 30

Leu Met Val Gly Trp Phe Arg Gin Ala Pro Gly Lys Glu Arg G1U Phe 35 40 45

Ala Ala Gly He Trp Ser Ser Gly Gly Thr Ala Tyr Ala Asp Ser Val 50 55 60

Lys Gly Arg Phe Thr Val Ser Arg Asp Asn Ala Lys Asn Thr Val Tyr 65 70 75 80

Leu Gin Met Asn Ser Leu Lys Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Gly Ser Tyr Asp Gly Asn Tyr Tyr lie Pro Gly Phe Tyr Lys Asp 100 105 110

Trp Gly Gin Gly Thr Gin Val Thr Val Ser Ser 115 120 <210> 15 <211> 123

<212> PRT <213> Lama glama <400> 15

Glu Val Gin Leu Val Glu Phe Gly Gly Gly Leu Val Gin Val Gly Asp 15 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Arg Thr Phe Ser Thr Tyr 20 25 30

Leu Met Val Gly Trp Phe Arg Gin Ala Pro Gly Lys Glu Arg Glu Phe 35 40 45

Ala Ala Gly lie Trp Ser Ser Gly Gly Thr Ala Tyr Ala Asp Ser Val 50 55 60

Lys Glu Arg Phe Thr Val Ser Arg Asp Asn Asp Lys Asn Thr Val Tyr 65 70 75 80

Leu Gin Met Asn Ser Leu Gin Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Gly Ser Tyr Asp Gly Asn Tyr Tyr lie Pro Gly Phe Tyr Lys Asp 100 105 110

Trp Gly Gin Gly Thr Gin Val Thr Val Ser Ser 115 120 <210> 16 <211> 123

<212> PRT <213> Lama glama <400> 16

Glu Val Gin Leu Val Glu Ser Gly Gly Gly Leu Val Gin Val Gly Asp 1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly His Thr Phe Ser Thr Tyr 20 25 30

Leu Met Val Gly Trp Phe Arg Gin Thr Pro Gly Lys Glu Arg Glu Phe 35 40 45

Ala Ala Val He Trp Ser Ser Gly Asp Thr Ala Tyr Ala Asp Ser Val 50 55 60

Lys Gly Arg Phe Thr He Ser Arg Asp Asn Ala Lys Asn Thr Val Tyr 65 70 75 80

Leu Gin Met Asn Ser Leu Lys Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Gly Ser Tyr Gly Gly Asn Tyr Tyr He Pro Gly Phe Tyr Glu Asp 100 105 110

Trp Gly Gin Gly Thr Gin Val Thr Val Ser Ser 115 120 <210> 17 <211> 126

<212> PRT <213> Lama glama <400> 17

Glu Val Gin Leu Val Glu Ser Gly Gly Gly Leu Val Gin Pro Gly Gly 15 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Ala Leu Asp Tyr Tyr zu zo ju

Ala lie Gly Trp Phe Arg Gin Val Pro Gly Lys Glu Arg Glu Gly Val 35 40 45

Ser Cys He Ser Ser Ser Asp Gly He Thr Tyr Tyr Val Asp Ser Val 50 55 60

Lys Gly Arg Phe Thr He Ser Arg Asp Asn Ala Lys Asn Thr Val Tyr 65 70 75 80

Leu Gin Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Thr Asp Ser Gly Gly Tyr He Asp Tyr Asp Cys Met Gly Leu Gly 100 105 110

Tyr Asp Tyr Trp Gly Gin Gly Thr Gin Val Thr Val Ser Ser 115 120 125 <210> 18 <211> 124

<212> PRT <213> Lama glama <400> 18

Glu Val Gin Leu Val Glu Ser Gly Gly Gly Leu Val Gin Ala Gly Gly 15 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Arg Thr Phe Ser Ser Tyr 20 25 30

Ala Met Gly Trp Phe Arg Gin Ala Pro Gly Lys Glu Leu Glu Phe Val 35 40 45

Thr Ala Val Ser Trp Ser Gly Gly Ser Thr Tyr Tyr Ala Asp Ser Val 50 55 60

Lys Gly Arg Phe Thr He Ser Arg Asp Ser Ala Lys Asn Thr Val Tyr 65 70 75 80

Leu Gin Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Ala Gin Ser Thr He Val Glu Val Thr Thr Leu Glu Ala Tyr Asp 100 105 110

Tyr Trp Gly Gin Gly Thr Gin Val Thr Val Ser Ser 115 120 <210> 19 <211> 122

<212> PRT <213> Lama glama <400> 19

Glu Val Gin Leu Val Glu Ser Gly Gly Gly Ser Val Gin Ala Gly Gly 1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Glu Arg Thr Phe Ser Pro Tyr 20 25 30

Ala Met Gly Trp Phe Arg Gin Ala Pro Gly Lys Glu Arg Glu Phe Val 35 40 45

Ala His lie Thr Trp Ser Ala Gly Ser Thr Tyr Tyr Ala Asp Ser Val 50 55 60

Lys Gly Arg Phe Thr He Ser Arg Asp Asn Ala Lys Asn Thr Val Tyr 65 70 75 80

Leu Gin Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Ala Lys Arg Arg Tyr Gly He Val Asp Arg Asp Tyr Asn Asp Trp 100 105 110

Gly Gin Gly Thr Gin Val Thr Val Ser Ser 115 120 <210> 20 <211> 116

<212> PRT <213> Lama glama <400> 20

Glu Val Gin Leu Val Glu Ser Gly Gly Gly Leu Val Gin Ala Gly Gly 15 10 15

Ser Leu Arg Leu Ala Cys He Ala Ser Gly Arg Asp He Phe Ser Val 20 25 30

Ser Ala Thr Gly Trp Tyr Arg Gin Ala Pro Gly Lys Gin Arg Glu Phe 35 40 45

Val Ala Gly He Ser Asn He Gly Ala Thr Lys Phe Ala Asp Ser Val 50 55 60

Lys Gly Arg Phe Thr lie Ser Arg Asp Asn Ala Lys Asn Thr Val Tyr 65 70 75 80

Leu Gin Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Asn Val Leu Leu Trp Ser Gly Asn Leu Trp Gly Gin Gly Thr Gin Val 100 105 110

Thr Val Ser Ser 115 <210> 21 <211> 116

<212> PRT <213> Lama glama <400> 21

Glu Val Gin Leu Val Glu Ser Gly Gly Gly Leu Val Gin Ala Gly Gly 15 10 15

Ser Leu Arg Leu Ser Cys lie Ala Ser Gly Arg Asp lie Phe Ser He 20 25 30

Thr Ala He Gly Trp Tyr Arg Gin Ala Pro Gly Lys Gin Arg Glu Phe 35 40 45

Val Ala Gly lie Ser Asn He Gly Ala Thr Lys Tyr Thr Asp Ser Val 50 55 60

Lys Gly Arg Phe Thr He Ser Gly Asp Asn Ala Glu Asn Thr Val Tyr 65 70 75 80

Leu Gin Met ASrt Ser Leu LyS Pro Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Asn Val Leu Leu Trp Ser Ala Asn Tyr Trp Gly Gin Qly Thr Gin Val 100 105 110

Thr Val Ser Ser 115 <210> 22 <211> 126

<212> PRT <213> Lama glama <400> 22

Glu Val Gin Leu Val Glu Ser Gly Gly Gly Leu Val Gin Ala Gly Gly 15 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Ala Phe Asp Asp His 20 25 30

Leu He Gly Trp Phe Arg Gin Ala Pro Gly Lys Glu Arg Glu Ala Val 35 40 45

Ser Cys He Ser Arg Ser Ala Gly Ser Thr Tyr Tyr Ala Asp Ser Val 50 55 60

Lys Gly Arg Phe Thr He Ser Gly Asp Asn Ala Lys Asn Thr Val Tyr 65 70 75 80

Leu Gin Met Asn Ser Leu Lys Ala Asp Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Ala Gly Pro Ala Trp Gly Arg Pro Ala Ser Pro Leu Pro Tyr Glu 100 105 110

Tyr Asp Tyr Trp Gly Gin Gly Thr Gin Val Thr Val Ser Ser 115 120 125 <210> 23 <211> 134

<212> PRT <213> Lama glama <400> 23

Glu Val Gin Leu Val Glu Ser Gly Gly Gly Leu Val Gin Ala Gly Gly 15 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Asp Asp Tyr 20 25 30

Ala lie Gly Trp Phe Arg Gin Ala Pro Gly Lys Glu Arg Glu Gly Val 35 40 45

Ser Cys He Ser Ser Ser Asp Gly Ser Thr Ala Tyr Ala Asp Ser Val 50 55 60

Lys Gly Arg Phe Thr He Ser Ser Asp Asn Ala Lys Asn Thr Val Tyr 65 70 75 80

Leu Gin Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Thr Ala Val Pro Ala Thr Arg Arg Thr Pro Gin Met Val Val Ala Asn 100 105 110

Val Cys Trp Leu Ala Glu Tyr Glu Tyr Asp Tyr Trp Gly Gin Gly Thr 115 120 125

Gin Val Thr Val Ser Ser 130 <210> 24 <211> 125

<212> PRT <213> Lama glama <400> 24

Glu Val Gin Leu Val Glu Ser Gly Gly Gly Leu Val Gin Ala Gly Gly 15 10 15

Ser Leu Arg Leu Ser Cys Val Ala Ser Gly Phe Thr Phe Asp Asp Tyr 20 25 30

Ala He Gly Trp Phe Arg Gin Ala Pro Gly Lys Glu Arg Glu Gly Val 35 40 45

Ser CyS He Ser Ser Ser Asp Gly Ser Thr Tyr Tyr Ala Asp Ser Val 50 55 60

Lys Gly Arg Phe Thr lie Ser Gly Asp Asn Ala Lys Asn Thr Val Tyr 65 70 75 80

Leu Gin Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Ala Gly He Thr Pro Cys Ser Asp Tyr Thr Gin Thr Tyr Glu Tyr 100 105 110

Asp Val Trp Gly Gin Gly Thr Gin Val Thr Val Ser Ser 115 120 125 <210> 25 <211> 134

<212> PRT <213> Lama glama <400> 25

Glu Val Gin Leu Val Glu Ser Gly Gly Gly Leu Val Gin Ala Gly Gly 15 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Asp Asp Tyr 20 25 30

Ala He Gly Trp Phe Arg Gin Ala Pro Gly Lys Glu Arg Glu Gly Val 35 40 45

Ser CyS He Ser Arg Ser Asp Gly Ser Thr Ser Tyr Ala Asp Ser Val 50 55 60

Lys Gly Arg Phe Thr He Ser Ser Asp Asn Ala Lys Asn Thr Val Tyr 65 70 75 80

Leu Val Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Thr Ala Val Pro Ala Thr Arg Arg Thr Pro Gin Met Val Val Ala Asn 100 105 110

Met Cys Trp Leu Ala Glu Tyr Glu Tyr Asp Tyr Trp Gly Gin Gly Thr 115 120 125

Gin Val Thr Val Ser Ser 130 <210> 26 <211> 125

<212> PRT <213> Lama glama <400> 26

Glu Val Gin Leu Val Glu Ser Gly Gly Gly Leu Val Gin Ala Gly Gly 15 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Asp Asp Tyr 20 25 30

Ala lie Gly Trp Phe Arg Gin Ala Pro Gly Lys Glu Arg Glu Gly Val 35 40 45

Ser Cys He Ser Ser Ser Asp Gly Ser Thr Tyr Tyr Ala Asp Ser Val 50 55 60

Lys Gly Arg Phe Thr He Ser Ser Asp Asn Ala Lys Asn Thr Val Tyr 65 70 75 80

Leu Gin Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Ala Gly He Thr Pro Cys Ser Asp Tyr Thr Gin Thr Tyr Glu Tyr 100 105 110

Asp Val Trp Gly Gin Gly Thr Gin Val Thr Val Ser Ser 115 120 125 <210> 27 <211> 134

<212> PRT <213> Lama glama <400> 27

Glu Val Gin Leu Val Glu Ser Gly Gly Gly Leu Val Gin Ala Gly Gly 1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Val Ser Gly Phe Thr Phe Asp Asp Tyr 20 25 30

Ala He Gly Trp Phe Arg Gin Ala Pro Gly Lys Glu Arg Glu Gly Val 35 40 45

Ser Cys He Ser Ser Ser Asp Gly Ser Thr Ala Tyr Ala Asp Ser Val 50 55 60

Lys Gly Arg Phe Thr He Ser Ser Asp Asn Ala Lys Asn Thr Val Tyr 65 70 75 80

Leu Gin Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Thr Ala Val Pro Ala Tht Arg Arg Thr Pro Gin Met Val Val Ala Asn 100 105 110

Val Cys Trp Leu Ala Glu Tyr Glu Tyr ASp Tyr Trp Gly Gin Gly Thr 115 120 125

Gin Val Thr Val Ser Ser 130 <210> 28 <211> 132

<212> PRT <213> Lama glama <400> 28

Glu Val Gin Leu Val Glu Ser Gly Gly Gly LeU Val Gin Ala Gly Gly 15 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Asp Asp Tyr 20 25 30

Ala He Gly Trp Phe Arg Gin Ala Pro Gly Lys Glu Arg Glu Gly Val 35 40 45

Ser Tyr He Ser Ser Ser Asp Gly Ser Thr Tyr Tyr Ala Asp Ser Val 50 55 60

Lys Gly Arg Phe Thr Ser Ser Ser Asp Asn Ala Lys Asn Thr Val Tyr 65 70 75 80

Leu Gin Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Ala Arg Pro Thr Leu Arg Val Arg Leu Asp Asn Asp Arg His His 100 105 110

Leu Leu Tyr Glu Tyr Glu Tyr Asp Tyr Trp Gly Gin Gly Thr Gin Val 115 120 125

Thr Val Ser Ser 130 <210> 29 <211> 134

<212> PRT <213> Lama glama <400> 29

Glu Val Gin Leu Val Glu Ser Gly Gly Gly Leu Val Gin Ala Gly Gly 15 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Asp Asp Tyr 20 25 30

Thr He Gly Trp Phe Arg Gin Ala Pro Gly Lys Glu Arg Glu Gly Val 35 40 45

Ser Cys lie Ser Ser Ser Asp Gly Ser Thr Ser Tyr Ala Asp Ser Val 50 55 60

Lys Gly Arg Phe Thr He Ser Ser Asp Asn Ala Lys Asn Thr Val Tyr 65 70 75 80

Leu Gin Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Thr Ala Val Pro Ala Thr Arg Arg Thr Pro Gin Met Val Val Leu Asn 100 105 110

Met Cys Trp Leu Ala Glu Tyr Glu Tyr Asp Tyr Trp Gly Gin Gly Thr 115 120 125

Gin Val Thr Val Ser Ser 130 <210> 30 <211> 125

<212> PRT <213> Lama glama <400> 30

Glu Val Gin Leu Val Glu Ser Gly Gly Gly Leu Val Gin Ala Gly Gly 15 10 15

Ser Leu Arg Leu Ser Cys Val Ala Ser Gly Phe Thr Phe Asp Asp Tyr 20 25 30

Ala lie Gly Trp Phe Arg Gin Ala Pro Gly Lys Glu Arg Glu Gly Val 35 40 45

Ser Cys He Ser Ser Ser Asp Gly Ser Thr Tyr Tyr Ala Asp Ser Val 50 55 60

Lys Gly Arg Phe Thr He Ser Set Asp Asn Ala Lys Asn Thr Val Tyr 65 70 75 80

Leu Gin Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Ala Gly He Thr Pro Cys Ser Asp Tyr Thr Gin Thr Tyr Glu Tyr 100 105 110

Asp Val Trp Gly Gin Gly Thr Gin Val Thr Val Ser Ser 115 120 125 <210> 31 <211> 134

<212> PRT <213> Lama glama <400> 31

Glu Val Gin Leu Val Glu Ser Gly Gly Gly Ser Val Gin Ala Gly Gly 15 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Asp Asp Tyr 20 25 30

Ala He Gly Trp Phe Arg Gin Ala Pro Gly Lys Glu Arg Glu Gly Val 35 40 45

Ser Cys lie Ser Ser Ser Asp Gly Ser Thr Asn Tyr Ala Asp Ser Val 50 55 60

Lys Gly Arg Phe Thr He Ser Ser Asp Thr Ala Lys Asn Thr Val Tyr 65 70 75 80

Leu Gin Met ASn Ser Leu Lys Pro Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Thr Ala Val Pro Ala Thr Arg Arg Thr Pro Gin Met Val Asp Ala Asn 100 105 HO

Met Cys Trp Leu Ala Glu Tyr Glu Tyr Asp Tyr Trp Gly Gin Gly Thr 115 120 125

Gin Val Thr Val Ser Ser 130 <210> 32 <211> 127

<212> PRT <213> Lama glama <400> 32

Glu Val Gin Leu Val Glu Ser Gly Gly Gly Leu Ala Gin Ala Gly Gly 1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Arg Arg Phe Gly Gly His 20 25 30

Ala Met Gly Trp Phe Arg Gin Ala Pro Gly Glu Glu Arg Glu Phe Val 35 40 45

Ala Thr He Tyr Trp Thr Ser Gly Met Thr Arg Tyr Ala Gly Ser Val 50 55 60

Lys Gly Arg Phe Thr He Ser Arg Asp Asn Ala Glu Asn Thr Val Phe 65 70 75 80

Leu Gin Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Val lie Lys Asp Phe Gin Leu Arg Val Asp Val Thr Ser Ala Ser 100 105 110

Ala Tyr Asp Tyr Trp Gly Gin Gly Thr Gin Val Thr Val Ser Ser 115 120 125 <210> 33 <211> 127

<212> PRT <213> Lama glama <400> 33

Gly Val Gin Leu Val Glu Ser Gly Gly Gly Leu Ala Gin Ala Gly Gly 1 5 10* 15* *

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Arg Arg Phe Gly Gly His 20 25 30

Ala Met Gly Trp Phe Arg Gin Ala Pro Gly Lys Asp Arg Glu Phe Val 35 40 45

Ala Thr lie Tyr Trp Thr Thr Gly Met Thr Arg Tyr Ala Asp Ser Val 50 55 60

Lys Gly Arg Phe Thr He Ser Arg Asp Asn Ala Glu Asn Thr Val Phe 65 70 75 80

Leu Gin Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Leu He Arg Asp Phe Asn He Arg Leu Asp Val Thr Ser Ala Ser 100 105 110

Ala Tyr Gly Tyr Trp Gly Gin Gly Thr Gin Val Thr Val Ser Ser 115 120 125 <210> 34 <211> 123

<212> PRT <213> Lama glama <400> 34

Glu Val Gin Leu Val Glu Ser Gly Gly Gly Leu Val Gin Val Gly Asp 15 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Arg Thr Phe Ser Thr Tyr 20 25 30

Leu Met Val Gly Trp Phe Arg Gin Ala Pro Gly Lys Glu Arg Glu Phe 35 40 45

Ala Ala Gly He Trp Ser Ser Gly Asp Thr Ala Tyr Ala Asp Ser Val 50 55 60

Arg Gly Arg Phe Thr He Ser Arg Asp Asn Ala Lys Asn Thr Val Tyr 65 70 75 80

Leu Gin Met Asn Ser Leu Lys Thr Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Gly Ser Tyr Asp Gly Asn Tyr Tyr He Pro Gly Phe Tyr Lys Asp 100 105 110

Trp Gly Gin Gly Thr Gin Val Thr Val Ser Ser 115 120 <210> 35 <211> 123

<212> PRT <213> Lama glama <400> 35

Glu Val Gin Leu Val Glu Ser Gly Gly Gly Leu Val Gin Val Gly Asp 15 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Arg Thr Phe Ser Thr Tyr 20 25 30

Leu Val Val Gly Trp Phe Arg Gin Ala Pro Gly Lys Glu Arg Glu Phe 35 40 45

Ala Ala Ala lie Trp Ser Ser Gly Asp Thr Ala Tyr Ala Asp Ser Val 50 55 60

Lys Gly Arg Phe Thr He Ser Arg Asp Asn Ala Lys Asn Thr Val Tyr 65 70 75 80

Leu Gin Met Asn Ser Leu Lys Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Gly Ser Tyr Gly Gly Asn Tyr Tyr He Pro Gly Phe Tyr Glu Asp 100 105 110

Trp Gly Gin Gly Thr Gin Val Thr Val Ser Ser 115 120 <210> 36 <211> 121

<212> PRT <213> Lama glama <400> 36

Glu Val Gin LeU Val Glu Ser Gly Gly Gly Leu Val Gin Ala Gly Gly 15 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Arg Thr Phe Ser Arg Asn 20 25 30

Ala Met Ala Trp Phe Arg Gin Val Pro Gly Lys Val Arg Glu Phe Val 35 40 45

Ala Gly He Arg Trp Asn Val Gly Arg Leu Asp Tyr Ala Asp Ser Val 50 55 60

Lys Gly Arg Phe Thr He Ser Arg Asp Asn Ala Glu Asn Thr Val Tyr 65 70 75 80

Leu Gin Met Asn Asp Leu Lys Thr Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Ala Tyr Ala Gly Leu Val Phe Ser Gly He Pro Asp Tyr Trp Gly 100 105 110

Gin Gly Thr Gin Val Thr Val Ser Ser 115 120 <210> 37 <211> 118

<212> PRT <213> Lama glama <400> 37

Glu Val Gin LeU Val Glu Ser Gly Arg Gly Leu Val Gin Ala Gly Gly 15 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Ser lie Phe Ser He Asn 20 25 30

Ala Met Leu Trp Tyr Arg Gin Ala Pro Gly Lys Gin Arg Glu Leu Val 35 40 45

Ala Ala He Thr Ser Gly Gly Ser Thr Ash Tyr Ala Asp Ser Val Lys 50 55 60

Gly Arg Phe Thr He Ser Arg Asp Asn Ala Lys Asn Thr Val Tyr Leu 65 70 75 80

Gin Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Val Tyr Tyr Cys Ala 85 90 95

Ala Asp Ser Asp Tyr Ser Ser Asp Tyr Tyr Tyr Trp Gly Gin Gly Thr 100 105 110

Gin Val Thr Val Ser Ser 115 <210> 38 <211> 133

<212> PRT <213> Lama glama <400> 38

Glu Val Gin Leu Val Glu Ser Gly Gly Gly Leu Val Gin Ser Gly Gly 15 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Ala Leu Asp Tyr Tyr 20 25 30

Thr He Gly Trp Phe Arg Glh Ala Pro Gly Lys Glu Arg Glu Gly Val 35 40 45

Ser Cys He Ser Gly Gly Asp Thr Ser Thr Tyr Tyr Ala Asp Ser Val 50 55 60

Lys Gly Arg Phe Thr He Ser Arg Asp Asn Ala Lys Asn Thr Val Tyr 65 70 75 80

Leu Gin Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Thr Asp Ser Ala Gly Val Pro Ala Gly Pro Ala Ala Val Tyr Gly 100 105 110

Ser Thr Cys Ser Arg Leu Glu Tyr Asp Tyr Trp Gly Gin Gly Ala Gin 115 120 125

Val Thr Val Ser Ser 130 <210> 39 <211> 124

<212> PRT <213> Lama glama <400> 39

Glu Val Gin Leu Val Glu Ser Gly Gly Gly Leu Val Gin Ala Gly Gly 15 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Gly Thr Phe Ser Ser Tyr 20 25 30

Ser Met Gly Trp Phe Arg Gin Ala Pro Gly Lys Glu Arg Glu Phe lie 35 40 45

Ala Ala He Asn Trp Asn Gly Asp Ser Thr Tyr Tyr Glu Asp Ser Val 50 55 60

Lys Gly Arg Phe Thr Val Ser Arg Asp Asn Ala Lys Asn Thr Val Tyr 65 70 75 80

Leu Gin Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Ala Thr Gly Trp Gly Arg Ala Tyr Glu Gin Ala Tyr Glu Tyr Asp 100 105 110

Val Trp Gly Gin Gly Thr Gin Val Thr Val Ser Ser 115 120 <210> 40 <211> 119

<212> PRT <213> Lama glama <400> 40

Glu Val Gin Leu Val Glu Ser Gly Gly Gly Leu Val Gin Pro Gly Gly 15 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Val Asp Asp Tyr 20 25 30

Ala Met Ser Trp Val Arg Gin Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45

Ser Thr lie Ser Trp Asn Asp Glu Tyr Thr Tyr Tyr Ala Glu Ser Met 50 55 60

Lys Gly Arg Phe Thr He Ser Arg Asp Asn Ala Lys Asn Thr Leu Tyr 65 70 75 80

Leu Gin Met Asn Ser Leu Lys Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Lys Gly Gly Ser Arg Leu Tyr Asp Tyr His Tyr Trp Gly Gin Gly 100 105 110

Thr Gin Val Thr Val Ser Ser 115 <210> 41 <211> 123

<212> PRT <213> Lama glama <400> 41

Glu val Gin Leu Val Glu Ser Gly Gly Gly Leu Val Gin Ala Gly Gly 15 10 15

Ser Leu Arg Leu Ser Cys Ala Ser Ser Gly Phe Thr Phe Asp Asp Tyr 20 25 30

Ala He Gly Trp Phe Arg Gin Ala Pro Gly Lys Glu Arg Glu Gly Val 35 40 45

Ser Cys He Ser Ser Ser Asp Gly Ser Thr Tyr Tyr Ala Asp Ser Val 50 55 60

Lys Gly Arg Phe Thr He Ser Ser Asp Asn Ala Lys Asn Thr Val Tyr 65 70 75 80

Leu Gin Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Ala Arg Pro Pro Phe His Ser Cys Ser Glu Tyr Glu Asn Asp Tyr 100 105 110

Trp Gly Gin Gly Thr Gin Val Thr Val Ser Ser 115 120 <210> 42 <211> 120

<212> PRT <213> Lama glama <400> 42 ijxu vai laxn heu vax wxu ser isxy isxy <sxy xeu vax isxn Axa isxy wxy 15 10 15

Ser Leu Arg lieu Ser Cys Ala Ala Ser Gly Arg Thr lie Ser Ser Ser 20 25 30

Val Leu Gly Trp Phe Arg Gin Ala Pro Gly Lys Glu Arg Glu Phe Val 35 40 45

Ala Ala He Ser Gly Ser Gly Ser Ser Thr Asp Ser Ala Lys Asp Arg 50 55 60

Phe Thr lie Ser Arg Asp Asn Ala Lys Asn Thr Val Tyr Leu Gin Met 65 70 75 80

Asn Ser Leu Lys Pro Glu Asp Thr Ala Val Tyr Tyr Cys Ala Ala Gly 85 90 95

Arg Ala Phe LeU Thr Arg Asp Pro Phe Tyr Tyr Asp He Trp Gly Gin 100 105 110

Gly Thr Gin Val Thr Val Ser Ser 115 120 <210> 43 <211> 116

<212> PRT <213> Lama glama <400> 43

Glu Val Gin Leu Val Glu Ser Gly Gly Gly Leu Val Gin Ala Gly Gly 15 10 15

Ser Leu Arg Leu Thr Cys lie Ala Ser Gly Arg Asp He Phe Ser Val 20 25 30

Thr Ala Met Gly Trp Tyr Arg Gin Ala Pro Gly Lys Gin Arg Glu Phe 35 40 45

Val Ala Gly Leu Ser Asn He Gly Ala Thr Lys Tyr Ala Asp Ser Val 50 55 60

Lys Gly Arg Phe Thr Val Ser Gly Asp Ala Ala Lys Asn Thr Val Tyr 65 70 75 80

Leu Gin Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Asn Val Leu Leu Trp Ser Gly Asn Tyr Trp Gly Gin Gly Thr Gin Val 100 105 110

Thr Val Ser Ser 115 <210> 44 <211> 132

<212> PRT <213> Lama glama <400> 44

Glu Val Gin Leu Val Glu Ser Gly Gly Gly Leu Val Gin Ala Gly Gly 15 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Asp Asp Tyr 20 25 30

Ala lie Gly Trp Phe Arg Gin Ala pro Gly Lys Glu Arg Glu Gly Val 35 40 45

Ser Tyr He Ser Lys Ser Asp Gly Thr Thr Tyr Tyr Ala Asp Ser Val 50 55 60

Lys Gly Arg Phe Thr Ser Ser Ser Asp Asn Ala Lys Asn Thr Val Tyr 65 70 75 80

Leu Gin He Asn Ser Leu Lys Pro Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Ala Arg Pro Thr Leu Arg Val Arg Leu Asp Asn Asp Arg His His 100 105 110

Leu Leu Tyr Glu Tyr Glu Tyr Asp Tyr Trp Gly Gin Gly Thr Gin Val 115 120 125

Thr Val Ser Ser 130 <210> 45 <211> 132

<212> PRT <213> Lama glama <400> 45

Glu Val Gin Leu Val Glu Ser Gly Gly Gly Leu Val Gin Ala Gly Gly 15 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Asp Glu Tyr 20 25 30

Ala He Gly Trp Phe Arg Gin Ala Pro Gly Lys Glu Arg Glu Gly Val 35 40 45

Ser Tyr lie Ser Ser Ser Asp Gly Ser Thr Tyr Tyr Ala Asp Ser Val 50 55 60

Lys Gly Arg Val Thr Ser Ser Ser Asp Asn Ala Lys Asn Thr Val Tyr 65 70 75 80

Leu Gin Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Ala Arg Pro Thr Leu Arg Val Arg Leu Asp Asn Asp Arg His His 100 105 110

Leu Leu Tyr Glu Tyr Glu Tyr Asp Tyr Trp Gly Gin Gly Thr Gin Val 115 120 125

Thr Val Ser Ser 130 <210> 46 <211> 124

<212> PRT <213> Lama glama <400> 46

Glu Val Gin Leu val Glu Ser Gly Gly Gly Leu Val Gin Ala Gly Gly 1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Arg Thr Phe Ser Ser Tyr 20 25 30

Ala Met Gly Trp Phe Arg Gin Ala Pro Gly Lys Glu Arg Glu Phe Val 35 40 45

Ala Arg He Ser Trp Asn Gly Gly Ser Thr Tyr His Ala Asp Ser Val 50 55 60

Lys Gly Arg Phe Thr He Ser Arg Asp Asn Pro Lys Asn Thr Val Tyr 65 70 75 80

Leu Gin Met Asp Ser Leu Lys Pro Glu Asp Ala Ala He Tyr Tyr Cys 85 90 95

Ala Ala Ser He Ala Leu Val Gly Gly Val Thr Pro His Ser Tyr Asp 100 105 110

Tyr Trp Gly Gin Gly Thr Gin Val Thr Val Ser Ser 115 120 <210> 47 <211> 123

<212> PRT <213> Lama glama <400> 47

Glu Val Gin Leu Val Glu Ser Gly Gly Gly Leu Val Gin Val Gly Asp 1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Arg Thr Phe Ser Thr Tyr 20 25 30

Leu Met Val Gly Trp Phe Arg Gin Ala Pro Gly Lys Glu Arg Glu Phe 35 40 45

Ala Ala Val lie Trp Ser Ser Gly Asp Thr Asp Tyr Ala Asp Ser Val 50 55 60

Lys Gly Arg Phe Thr He Ser Arg Asp Asn Ala Lys Asn Thr Val Tyr 65 70 75 80

Leu Gin Met Asn Ser Leu Lys Ala Glu Asp Thr Ala Val Tyr Tyr CyS 85 90 95

Ala Gly Ser Tyr Gly Gly Asn Tyr Tyr He Pro Gly Phe Tyr Lys Asp 100 105 110

Trp Gly Gin Gly Thr Gin Val Thr Val Ser Ser 115 120 <210> 48 <211> 123

<212> PRT <213> Lama glama <400> 48

Glu Val Gin Leu Val Glu Ser Gly Gly Gly Leu Val Gin Val Gly Asp 15 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Arg Thr Phe Ser Thr Tyr 20 25 30

Leu Met Val Gly Trp Phe Arg Gin Ala Pro Gly Lys Glu Arg Glu Phe 35 40 45

Ala Ala Gly He Trp Ser Ser Gly Gly Thr Ala Tyr Ala Asp Ser Val 50 55 60

Lys Gly Arg Phe Thr lie Ser Arg Asp Asn Ala Lys Asn Thr Val Tyr 65 70 75 80

Leu Gin Met Asn Ser Leu Lys Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Gly Ser Tyr Asp Gly Asn Tyr Tyr He Pro Gly Phe Tyr Lys Asp 100 105 110

Trp Gly Gin Gly Thr Gin Val Thr Val Ser Ser 115 120 <210> 49 <211> 128

<212> PRT <213> Lama glama <400> 49

Glu Val Gin Leu Val Glu Ser Gly Gly Gly Leu Val Gin Ala Gly Gly 15 10 15

Ser Leu Gly Leu Ser Cys Ala Ala Ser Glu Arg Thr Leu Pro Ser Tyr 20 25 30

Val Met Gly Trp phe Arg Gin Ala Pro Gly Lys Glu Leu Glu Phe Val 35 40 45

Ala Gly lie Ser Trp Ser Ser Gly Arg Thr Tyr Tyr Thr Asp Ser Val 50 55 60

Lys Gly Arg Phe Thr He Ser Arg Asp Ala Ala Glu Asn Thr Trp Tyr 65 70 75 80

Leu Gin Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Ser Asn Ser Val Ser Glu Pro Thr Leu His Thr Trp Gin Tyr Glu 100 105 110

Ala Ser Tyr Asp Tyr Trp Gly Gin Gly Thr Gin Val Thr Val Ser Ser 115 120 125 <210> 50 <211> 124

<212> PRT <213> Lama glama <400> 50

Glu Val Gin Leu Val Glu Ser Gly Gly Gly Leu Val Gin Ala Gly Gly 15 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly His Thr Phe Ser Arg Tyr 20 25 30

Ala Met Gly Trp Phe Arg Arg Val Pro Gly Glu Glu Arg Glu Phe Val 35 40 45

Thr His He Thr Trp Asn Arg Gly Ser Thr Tyr Tyr Ala Asp Ser Val 50 55 60

Lys Gly Arg Phe Thr He Ser Arg Asp Lys Ala Ser ASn Thr Leu Tyr 65 70 75 80

Leu Gin Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Ala Gin He Lys Tyr Gly Glu He Thr His Pro Glu Glu Tyr Ser 100 105 110

Tyr Trp Gly Gin Gly Thr Gin Val Thr Val Ser Ser 115 120 <210> 51 <211> 121

<212> PRT <213> Lama glama <400> 51

Glu Val Gin Leu Val Glu Ser Gly Gly Gly Leu Val Gin Ala Gly Gly 15 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Arg Thr Phe Ser Arg Asn 20 25 30

Ala Met Gly Trp Phe Arg Gin Val Pro Gly Lys Ala Arg Glu Phe Val 35 40 45

Ala Ala He Arg Trp Asn Val Gly Arg Leu Asp Tyr Ala Asp Ser Val 50 55 60

Lys Gly Arg Phe Ala lie Ser Arg Asp Asn Ala Glu Asn Thr Val Tyr 65 70 75 80

Leu Gin Met Asn Asp Leu Lys Thr Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Ala Tyr Ala Gly Leu Val Tyr Ser Gly He Pro Asp Tyr Trp Gly 100 105 110

Gin Gly Thr Gin Val Thr Val Ser Ser 115 120 <210> 52 <211> 124

<212> PRT <213> Lama glama <400> 52

Glu Val Gin Leu Val Glu Ser Gly Gly Gly Leu Val Gin Ala Gly Gly 15 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly His Thr Phe Ser Arg Tyr 20 25 30

Ala Met Gly Trp Phe Arg Arg Val Pro Gly Lys Glu Arg Glu Phe Val 35 40 45

Thr His He Thr Trp Asn Arg Gly Ser Thr Tyr Tyr Ala Asp Ser Val 50 55 60

Lys Gly Arg Phe Thr He Ser Arg Asp Lys Ala Ser Asn Thr Leu Tyr 65 70 75 80

Leu Gin Met Asn Ser Leu Lys Pro G1U Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Ala Gin He Lys Tyt Gly Glu He Thr His Pro Glu Glu Tyr Ser 100 105 110

Tyr Trp Gly Gin Gly Thr Gin Val Thr Val Ser Ser 115 120 <210> 53 <211> 124

<212> PRT <213> Lama glama <400> 53

Glu Val Gin Leu Val Glu Ser Gly Gly Gly Leu Val Gin Ala Gly Asp 15 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly His Thr Phe Ser Arg Tyr 20 25 30

Ala Met Gly Trp Phe Arg Arg Val Pro Gly Lys Glu Arg Glu Phe Val 35 40 45

Thr His lie Thr Trp Asn Arg Gly Ser Thr Tyr Tyr Ala Asp Ser Val 50 55 60

Lys Gly Arg Phe Thr He Ser Arg Asp Lys Ala Ser Asn Thr Leu Tyr 65 70 75 80

Leu Gin Met Asn Ser Leu Lys Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Ala Gin Thr Lys Tyr Gly Glu He Thr Arg Pro Glu Glu Tyr Ser 100 105 110

Tyr Trp Gly Gin Gly Thr Gin Val Thr Val Ser Ser 115 120 <210> 54 <211> 115

<212> PRT <213> Lama glama <400> 54

Glu Val Gin Leu Val Glu Ser Gly Gly Gly Leu Val Gin Ala Gly Gly 15 10 15

Ser Leu Arg Leu Val Cys Ser Ala Ser Gly He Asp Phe Ser He Asn 20 25 30

Ala Met Ala Trp Tyr Arg Gin Ala Pro Gly Lys Gin Arg Glu Trp Val 35 40 45

Ala Phe Met He Asn Asp Ser Ser Thr Asp Tyr Thr Asp Ser Val Lys 50 55 60

Gly Arg Phe Thr He Ser Arg Asp Ser Thr Lys Asn He Leu Tyr Leu 65 70 75 80

Gin Met Asn Asn Leu Asn Val Glu Asp Thr Ala Val Tyr Tyr Cys Asn 85 90 95

Thr Ala Tyr Glu Gin His Thr Tyr Trp Gly Gin Gly Thr Gin Val Thr 100 105 110

Val Ser Ser 115 <210> 55 <211> 128

<212> PRT <213> Lama glama <400> 55

Glu Val Gin Leu Val Glu Ser Gly Gly Gly Leu Val Gin Ala Gly Gly 1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Asp Asp Tyr 20 25 30

Ala lie Ala Trp Phe Arg Gin Ala Pro Gly Lys Glu Arg Glu Gly He 35 40 45

Ser Cys He Thr Pro Ser Asp Asp Arg Thr Tyr Tyr Ala Asp Ser Val 50 55 60

Lys Gly Arg Phe He lie Ser Ser Asp Asn Ala Lys Asn Thr Val Tyr 65 70 75 80

Leu Gin Met Asn Ser Leu Lys Pro GlU Asp Thr Ala Asp Tyr Tyr Cys 85 90 95

Ala Ala Val Pro Arg Leu Arg Gly Leu Gly Tyr Trp Pro Tyr Pro Glu 100 105 110

Tyr Glu Tyr Asp Tyr Trp Gly Gin Gly Thr Gin Val Thr Val Ser Ser 115 120 125 <210> 56 <211> 124

<212> PRT <213> Lama glama <400> 56

Lys Val Gin LeU Val Glu Ser Gly Gly Gly Leu Val Gin Ala Gly Gly 15 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Asp Asp Tyr 20 25 30

Ala Leu Gly Trp Phe Arg Gin Ala Ala Gly Lys Glu Arg Glu Gly Val 35 40 45

Ser Cys He Arg Cys Ser Asp Gly Ser Thr Tyr Tyr Ala ASp Ser Val 50 55 60

Lys Gly Arg Phe Thr lie Ser Ser Asp Asn Ala Lys Asn Thr Val Tyr 65 70 75 80

Leu Gin Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Ala Ser He Val Pro Arg Ser Lys Leu Glu Pro Tyr Glu Tyr Asp 100 105 110

Ala Trp Gly Gin Gly Thr Gin Val Thr Val Ser Ser 115 120 <210> 57 <211> 119

<212> PRT <213> Lama glama <400> 57

Glu Val Gin Leu Val Glu Ser Gly Gly Gly Leu Val Gin Pro Gly Gly 15 10 15

Ser Leu Arg Leu Ser Cys Ala Val Ser Gly Phe Thr Val Asp Asp Tyr 20 25 30

Ala Met Ser Ttp Val Arg Gin Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45

Ser Thr He Ser Trp Asn Asp Glu Tyr Thr Tyr Tyr Ala Glu Ser Met 50 55 60

Lys Gly Arg Phe Thr He Ser Arg Asp Asn Ala Lys Asn Thr Leu Tyr 65 70 75 80

Leu Gin Met Asn Ser Leu Lys Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Lys Gly Gly Ser Arg Leu Tyr Asp Tyr His Tyr Trp Gly Gin Gly 100 105 110

Thr Gin Val Thr Val Ser Ser 115 <210> 58 <211> 131

<212> PRT <213> Lama glama <400> 58

Glu Val Gin Leu Val Glu Ser Gly Gly Gly Leu Val Gin Ala Gly Gly 1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Leu Asp Asp Tyr 20 25 30

Ala lie Gly Trp Phe Arg Gin Ala Pro Gly Lys Glu Arg Glu Gly Val 35 40 45

Ser Cys lie Ser Ser Ser Asp Gly Ser Thr Thr Tyr Ala Asp Ser Val 50 55 60

Lys Gly Arg Phe Thr Val Ser Ser Asp Asn Ala Lys Asn Thr Val Tyr 65 70 75 80

Leu Gin Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Ala Gly Leu Arg Gly Arg Tyr Tyr Arg Gly Thr Tyr Ser Leu Val 100 105 110

Cys Ala Pro Tyr Glu Tyr Asp Phe Trp Gly Gin Gly Thr Gin Val Thr 115 120 125

Val Ser Ser 130 <210> 59 <211> 131

<212> PRT <213> Lama glama <400> 59

Glu Val Gin Leu Val Glu Ser Gly Gly Gly Leu Val Gin Ala Gly Gly 15 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Leu Asp Asp Tyr 20 25 30

Ala He Gly Trp Phe Arg Gin Ala Pro Gly Lys G1U Atg Glu Gly Val 35 40 45

Ser Cys He Ser Ser Ser Asp Gly Ser Thr Tyr Tyr Ala Asp Ser Val 50 55 60

Lys Gly Arg Phe Thr He Ser Ala Asp Asn Ala Lys Asn Thr Val Tyr 65 70 75 80

Leu Gin Met His Ser Leu Lys Pro Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Ala Gly Leu Arg Gly Arg Tyr Tyr Ser Gly Ser Asn Tyr Leu Val 100 105 110

Cys Ala Pro Tyr Glu Tyr Asp Tyr Trp Gly Gin Gly Thr Gin Val Thr 115 120 125

Val Ser Ser 130 <210> 60 <211> 125

<212> PRT <213> Lama glama <400> 60

Glu Val Gin Leu Val Glu Ser Gly Gly Gly Leu Val Gin Ala Gly Gly 15 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Asp Asp Tyr 20 25 30

Ala lie Gly Trp Phe Arg Gin Ala Pro Gly Lys Glu Arg Glu Gly Val 35 40 45

Ser Cys He Ser Ser Ser Asp Gly Ser Thr Tyr Tyr Ala Asp Ser Val 50 55 60

Lys Gly Arg Phe Thr He Ser Ser Asp Asn Ala Lys Asn Thr Val Tyr

65 70 75 BO

Leu Gin Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Ala Asp Phe He He Ser Ser Lys Arg Leu Cys Leu Asp Leu Phe 100 105 110

Gly Ser Arg Gly Gin Gly Thr Gin Val Thr Val Ser Ser 115 120 125 <210> 61 <211> 125

<212> PRT <213> Lama glama <400> 61

Glu Val Gin Leu Val Glu Ser Gly Gly Gly Ser Val Gin Ala Gly Gly 15 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Asp ASp Tyr 20 25 30

Ala He Gly Trp Phe Arg Gin Ala Pro Gly Lys Glu Arg Glu Gly Val 35 40 45

Ser Cys He Ser Ser Ser Asp Gly Ser Thr Tyr Tyr Ala Asp Ser Val 50 55 60

Lys Gly Arg Phe Thr He Ser Ser Asp Asn Ala Lys Asn Thr val Tyr 65 70 75 80

Leu Gin Met Asn Ser Leu Lys Pro Gly Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Ala Gly He Thr Pro Cys Ser Asp Tyr Thr Gin Thr Tyr Glu Tyr 100 105 110

Asp Val Trp Gly Gin Gly Thr Gin Val Thr Val Ser Ser 115 120 125 <210> 62 <211> 134

<212> PRT <213> Lama glama <400> 62

Glu Val Gin Leu Val Glu Ser Gly Gly Gly Leu Val Gin Ala Gly Gly 1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Asp Asp Tyr 20 25 30

Ala lie Gly Trp Phe Arg Arg Ala Pro Gly Lys Glu Arg Glu Gly Val 35 40 45

Ser Cys He Thr Ser Ser Asp Gly Ser Thr Ser Tyr Ala Asp Ser Val 50 55 60

Lys Gly Arg Phe Thr He Ser Ser Asp Ser Ala Lys Asn Thr Val Tyr 65 70 75 80

Leu Gin Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Ser Val Val Pro Ala Thr Arg Arg Asn Pro Gin Met Val Val Ala Lys 100 105 110

Lys Cys Trp Leu Ala Glu Tyr Glu Tyr Asp Tyr Trp Gly Gin Gly Thr 115 120 125

Gin Val Thr Val Ser Ser 130 <210> 63 <211> 130

<212> PRT <213> Lama glama <400> 63

Glu Val Gin Leu Val Glu Ser Gly Gly Gly Leu Val Gin Ala Gly Gly 15 10 15

Ser Leu Arg LeU Ser Cys Ala Val Ser Gly Phe Thr Phe Asp Asp Tyr 20 25 30

Ala He Gly Trp Phe Arg Gin Ala Pro Gly Lys Glu Arg Glu Gly Val 35 40 45

Ser Cys He Ser Arg Ser Asp Gly Ser Pro Tyr Tyr Ala Asp Ser Val 50 55 60

Lys Gly Arg Phe Thr lie Ser Ser Asp Asn Ala Lys Gly Thr Val Tyr 65 70 75 80

Leu Gin Met Ser Ser Leu Lys Pro Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Ala Ser Trp Ser Gly Ala Tyr Tyr Ser Gly Thr Tyr Tyr Cys Asp 100 105 110

Arg Leu Tyr Glu Tyr Asp Ala Trp Gly Gin Gly Thr Gin Val Thr Val 115 120 125

Ser Ser 130 <210> 64 <211> 123

<212> PRT <213> Lama glama <400> 64

Glu Val Gin Leu Val Glu Ser Gly Gly Gly Leu Val Gin Val Gly Asp 15 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Glu Arg Thr Phe Ser Thr Tyr 20 25 30

Leu Met Val Gly Trp Phe Arg Gin Ala Pro Gly Lys Glu Arg Glu Phe 35 40 45

Ala Ala Thr Met Trp Ser Ser Gly Asp Thr Ala Tyr Ala Asp Ser Val 50 55 60

Lys Gly Arg Phe Thr He Ser Arg Asp Asn Ala Lys Asn Thr Val Tyr 65 70 75 80

Leu Gin Met Asn Ser Leu Lys Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Gly Ser Tyr Gly Gly Asn Tyr Tyr He Pro Gly Phe Tyr Glu Asp 100 105 110

Trp Gly Gin Gly Thr Gin Val Thr Val Ser Ser 115 120 <210> 65 <211> 127

<212> PRT <213> Lama glama <400> 65

Glu Val Gin Leu Val Glu Ser Gly Gly Gly Leu Val Gin Pro Gly Gly 15 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Ala Leu Asp Tyr Tyr 20 - 25 30

Ala lie Gly Trp Phe Arg Gin Ala Pro Gly Lys Glu Arg Glu Gly Val 35 40 45

Ser Cys He Ser Ser Ser Asp Gly Ser Thr Tyr Tyr Ala Asp Ser Val 50 55 60

Lys Gly Arg Phe Thr He Ser Arg Asp Asn Ala Lys Asn Thr Val Tyr 65 70 75 80

Leu Gin Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Thr Asp Ser Leu Gly Tyr Gly Ser Ser Cys Arg Met Ala Pro Tyr 100 105 110

Glu Tyr Asp Tyr Trp Gly Gin Gly Thr Gin Val Thr Val Ser Ser 115 120 125 <210> 66 <211> 123

<212> PRT <213> Lama glama <400> 66

Glu Val Gin Leu Val Glu Ser Gly Gly Gly Leu Val Gin Val Gly Asp 15 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Arg Thr Phe Ser Thr Tyr 20 25 30

Leu Met Val Gly Trp Phe Arg Gin Ala Pro Gly Lys Glu Arg Glu Phe 35 40 45

Ala Ala Val He Trp Ser Ser Gly Gly Thr Ala Tyr Ala Asp Ser Val 50 55 60

Lys Gly Arg Phe Thr He Ser Arg Asp Asn Ala Lys Asn Thr Val Tyr 65 70 75 80

Leu Gin Met Asn Ser Leu Lys Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Gly Ser Tyr Asp Gly Asn Tyr Tyr He Pro Gly Phe Tyr Lys Asp 100 105 110

Trp Gly Gin Gly Thr Gin Val Thr Val Ser Ser 115 120 <210> 67 <211> 130

<212> PRT <213> Lama glama <400> 67

Glu Val Gin Leu Val Glu Ser Gly Gly Gly Leu Val Gin Pro Gly Gly 15 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Ser Asp Asp Tyr 20 25 30

Ala He Gly Trp Phe Arg Gin Ala Pro Gly Lys Glu Arg Glu Gly Val 35 40 45

Ser Cys He Ser Ser Ser Asp Gly He Thr Tyr Tyr Ala Asp Ser Val 50 55 60

Lys Gly Arg Phe Thr He Ser Arg Asp Ash Ala Lys Asn Thr Val Tyr 65 70 75 80

Leu Gin Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Thr Gly Asn Arg Arg lie Tyr Tyr Ser Asp Tyr Ala Leu Ala Cys 100 105 110

Phe Pro Tyr Glu Tyr Asp Tyr Trp Gly Gin Gly Thr Gin Val Thr Val 115 120 125

Ser Ser 130 <210> 68 <211> 123

<212> PRT <213> Lama glama <400> 68

Glu Val Gin Leu Val Glu Ser Gly Arg Arg Leu Val Gin Val Gly Asp 15 10 15

Ser Leu Arg Leu Ala Cys Ala Ala Ser Gly Arg Thr Phe Ser Thr Tyr 20 25 30

Leu Met Val Gly Trp Phe Arg Gin Ala Pro Gly Lys Glu Arg Glu Phe 35 40 45

Ala Ala Gly He Trp Ser Ser Gly Asp Thr Ala Tyr Ala Asp Ser Val 50 55 60

Lys Gly Arg Phe Thr He Ser Arg Asp Asn Ala Lys Asn Thr Val Tyr 65 70 75 80

Leu Gin Met Asn Ser Leu Lys Thr Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Gly Ser Tyr Asp Gly Asn Tyr Tyr He Pro Gly Phe Tyr Lys Asp 100 105 110

Trp Gly Gin Gly Thr Gin Val Thr Val Ser Sen 115 120 <210> 69 <211> 123

<212> PRT <213> Lama glama <400> 69

Glu Val Gin Leu Val Glu Ser Gly Gly Gly Leu Val Gin Val Gly Asp 1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Glu Arg Thr Phe Ser Thr Tyr 20 25 30

Leu Met Val Gly Trp Phe Arg Gin Ala Pro Gly Lys Glu Arg Glu Phe 35 40 45

Ala Ala Thr Met Trp Val Ser Gly Asp Thr Ala Tyr Ala Asp Ser Val 50 55 60

Lys Gly Arg Phe Thr lie Ser Arg Asp Asn Pro Lys Asn Thr Val Tyr 65 70 75 80

Leu Gin Met Asn Ser Leu Lys Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Gly Ser Tyr Gly Gly Asn Tyr Tyr He Pro Gly Phe Tyr LyS Asp 100 105 110

Trp Gly Gin Gly Thr Gin Val Thr Val Ser Ser 115 120 <210> 70 <211> 123

<212> PRT <213> Lama glama <400> 70

Glu Val Gin Leu Val Glu Ser Gly Gly Gly Leu Val Gin lie Gly Asp 1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Arg Thr Phe Ser Thr Tyr 20 25 30

Leu Met Val Gly Trp Phe Arg Gin Ala Pro Gly LyS Glu Arg GlU Phe 35 40 45

Ala Ala Ala He Trp Ser Ser Gly Asp Thr Ala Val Ala Asp Ser Val 50 55 60

Lys Gly Arg Phe Thr lie Ser Arg Asp Asn Ala Lys Asn Thr Val Tyr 65 70 75 80

Leu Gin Met Asn Ser Leu Lys Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Gly Ser Tyr Gly Gly Asn Tyr Tyr lie Pro Gly Phe Tyr Glu Asp 100 105 110

Trp Gly Gin Gly Thr Gin Val Thr Val Ser Ser 115 120 <210> 71 <211> 122

<212> PRT <213> Lama glama <400> 71

Glu Val Gin Leu Val Glu Ser Gly Gly Gly Leu Val Gin Pro Gly Gly 15 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Leu Asp Tyr Tyr 20 25 30

Ala He Gly Trp Phe Arg Gin Ala Pro Gly Lys Glu Trp Glu Gly Val 35 40 45

Ser Cys He Ser Ser Ser Asp Gly Ser Thr Tyr Tyr Ala Asp Ser Val 50 55 60

Lys Gly Arg Phe Thr He Ser Arg Asp Asn Ala Lys Asn Thr Val Tyr 65 70 75 80

Leu Gin Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Thr Asp Ser lie Val Cys Gly Ser Tyr Tyr Gly Met Asp Tyr Trp 100 105 110

Gly Lys Gly Thr Gin Val Thr Val Set Ser 115 120 <210> 72 <211> 123

<212> PRT <213> Lama glama <400> 72

Glu Val Gin Leu Val Glu Ser Gly Gly Ala Ser Val Gin Pro Gly Gly 15 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Arg Thr Phe Ser Thr Tyr 20 25 30

Leu Met Val Gly Trp Phe Arg Gin Ala Pro Gly Lys Glu Arg Glu Phe 35 40 45

Ala Ala Gly He Trp Ser Ser Gly Gly Thr Ala Tyr Ala Asp Ser Val 50 55 60

Lys Glu Arg Phe Thr Val Ser Arg Asp Asn Asp Lys Asn Thr Val Tyr 65 70 75 80

Leu Gin Met Asn Ser Leu Gin Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Gly Ser Tyr Asp Gly Asn Tyr Tyr lie Pro Gly Phe Tyr Lys Asp 100 105 110

Trp Gly Gin Gly Thr Gin Val Thr Val Ser Ser 115 120 <210> 73 <211> 128

<212> PRT <213> Lama glama <400> 73

Glu val Gin Leu val Glu Ser Gly Gly Gly Leu Val Gin Ala Gly Gly 1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Asp Asp Tyr 20 25 30

Ala lie Gly Trp Phe Arg Gin Ala Pro Gly Lys Glu Arg Glu Gly Val 35 40 45

Ser Cys He Ser Ser Ser ASp Gly Ser Thr Tyr Tyr Ala Asp Ser Val 50 55 60

Lys Gly Arg Phe Thr He Ser Ser Asp Asn Ala Lys Asn Thr Val Tyr 65 70 75 80

Leu Gin Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Ala Ala Trp Gly Ala Ser Arg Leu Pro He Gly Thr Met Pro Pro 100 105 110

Tyr Glu Tyr Asp Tyr Trp Gly Gin Gly Thr Gin Val Thr Val Ser Ser 115 120 125 <210> 74 <211> 118

<212> PRT <213> Lama glama <400> 74

Glu Val Gin Leu Val Glu Ser Gly Gly Gly Leu Val Gin Ala Gly Gly 15 10 15

Ser Leu Arg Leu Val Cys Ser Ala Ser Gly He Asp Phe Ser He Asn 20 25 30

Val Met Gly Trp Tyr Arg Gin Ala Pro Gly Lys Gin Arg Glu Trp Val 35 40 45

Ala Phe lie Gly Ser Gly Gly Ser Thr Asp Tyr lie Asp Tyr Thr Asp 50 55 60

Ser Val Lys Gly Arg Phe Thr He Ser Arg Asp Ser Thr Lys Asn He 65 70 75 80

Leu Tyr Leu Arg Met Asn Asn Leu Asn Val Glu Asp Thr Ala Val Tyr 85 90 95

Tyr Cys Asn Thr Ala Tyr Glu Gin His Thr Tyr Trp Gly Gin Gly Thr 100 105 110

Gin Val Thr Val Ser Ser 115 <210> 75 <211> 125

<212> PRT <213> Lama glama <400> 75

Glu Val Gin Leu Val Glu Ser Gly Gly Gly Leu Val Gin Ala Gly Gly 1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Asp Asp Tyr 20 25 30

Ala He Gly Trp Phe Arg Gin Ala Pro Gly LyS Asp Leu Glu Gly Val 35 40 45

Ser Cys He Ser Ser Ser Asp Gly Ser Thr Tyr Tyr Ala Asp Ser Val 50 55 60

Lys Gly Arg Phe Thr lie Ser Ser Asp Asn Ala Lys Asn Thr Val Tyr 65 70 75 80

Leu Gin Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Ala Val Asn Gly Leu Gly Pro Phe Ser Val Pro Val Pro Val Tyr 100 105 110

Asp Phe Trp Gly Gin Gly Thr Gin Val Thr Val Ser Ser 115 120 125 <210> 76 <211> 130

<212> PRT <213> Lama glama <400> 76

Glu Val Gin Leu Val Glu Ser Gly Gly Gly Leu Val Gin Ala Gly Gly 15 10 15

Ser Leu Arg Leu Ser Gys Ala Val Ser Gly Leu Pro Phe Asp Asp Tyr 20 25 30

Val lie Gly Trp Phe Arg Gin Ala Pro Gly Lys Glu Arg Glu Gly Val 35 40 45

Ser Cys lie Ser Ser Ser Asp Gly Ser Thr Tyr Tyr Ala Asp Ser Val 50 55 60

Lys Gly Arg Phe Thr lie Ser Ser Asp Asn Ala Lys Asn Thr Val Tyr 65 70 75 80

Leu Gin Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Ala Tyr Tyr Cys 85 90 95

Ala Ala Gly Gly Pro Arg He Asn He Ala Thr Met Thr Cys Ser His 100 105 110

Asp Glu Tyr Glu Tyr Asp Tyr Trp Gly Gin Gly Thr Gin Val Thr Val 115 120 125

Ser Ser 130 <210> 77 <211> 133

<212> PRT <213> Lama glama <400> 77

Glu Val Gin LeU Val Glu Ser Gly Gly Gly Leu Val Gin Pro Gly Gly 15 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Ala Leu Asp Tyr Tyr 20 25 30

Ala He Gly Trp Phe Arg Gin Ala Pro Gly Lys Glu Arg Glu Gly Val 35 40 45

Ser Cys He Glu Ser Ser Asp Gly Ser Thr Tyr Tyr Ala Asp Ser Val 50 55 60

Lys Gly Arg Phe Thr He Ser Arg Asp Asn Ala Lys Asn Thr Val Tyr 65 70 75 80

Leu Gin Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Thr Asp Ser Ala Gly Val Pro Ala Gly Pro Ala Ala Val Tyr Gly 100 105 110

Ser Ser Cys Ser Arg Leu Glu Tyr Asp Tyr Trp Gly Gin Gly Thr Gin 115 120 125

Val Thr Val Ser Ser 130 <210> 78 <211> 124

<212> PRT <213> Lama glama <400> 78

Glu Val Gin Leu Val Glu Ser Gly Gly Gly Leu Val Gin Pro Gly Gly 15 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Gly Ser Tyr 20 25 30

Asp Met Ser Trp Val Arg Gin Ala Pro Gly Lys Gly Pro Glu Trp Val 35 40 45

Ser Ala lie Asn Ser Arg Gly Gly Ser Thr Tyr Tyr Ala Asp Ser Val 50 55 60

Lys Gly Arg Phe Thr He Ser Arg Asp Asn Ala Lys Asn Thr Leu Tyr 65 70 75 80

Leu Gin Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Thr Asp Pro Tyr Ser Leu Ser Tyr Tyr Gly Tyr Pro Leu Tyr Asp 100 105 110

Tyr Trp Gly Gin Gly Thr Gin Val Thr Val Ser Ser 115 120 <210> 79 <211> 115

<212> PRT <213> Lama glama <400> 79

Glu val Gin Leu Val Glu Ser Gly Gly Gly Leu Val Gin Ala Gly Gly 15 10 15

Ser Leu Arg Leu Val Cys Ser Ala Ser Gly He Asp Phe Ser lie Asn 20 25 30

Val Met Gly Trp Tyr Arg Gin Ala Pro Gly Lys Gin Arg Glu Trp Val 35 40 45

Ala Phe He Gly Ser Gly Ser Ser Thr Gly Tyr Thr Asp Ser Val Lys 50 55 60

Gly Arg Phe Ser He Ser Arg Asp Ser Thr Lys Asn He Leu Tyr Leu C5 7Π TR ΒΠ

Gin Met Asn Asn Leu Asn Val Glu Asp Thr Ala Val Tyr Tyr Cys Asn 85 90 95

Thr Ala Tyr Glu Gin His Thr Tyr Trp Gly Gin Gly Thr Gin Val Thr 100 105 110

Val Ser Ser 115 <210> 80 <211> 129

<212> PRT <213> Lama glama <400> 80

Glu Val Gin Leu Val Glu Ser Gly Gly Gly Leu Val Gin Ala Gly Gly 15 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Leu Asp Asp Tyr 20 25 30

Ala lie Gly Trp Phe Arg Gin Ala Pro Gly Lys Glu Arg Glu Gly Val 35 40 45

Ser Cys He Arg Asp Ser Asp Gly Ser Thr Tyr Tyr Ala Asp Ser Val 50 55 60

Lys Gly Arg Phe Thr He Ser Ser Asp Asn Asp Lys Asn Thr val Tyr 65 70 75 80

Leu Gin Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Ala Val Pro Ala Gly Arg Leu Arg Phe Gly Glu Gin Trp Tyr Pro 100 105 110

Leu Tyr Glu Tyr Asp Ala Trp Gly Gin Gly Thr Gin val Thr Val Ser 115 120 125

Ser <210> 81 <211> 124

<212> PRT <213> Lama glama <400> 81

Glu Val Gin Leu Val Glu Ser Gly Gly Gly Val Val Gin Ala Gly Asp 15 10 15

Ser Val Arg Leu Ser CyS Ala Ala Ser Gly Pro Thr Phe Arg Ser Tyr 20 25 30

Thr Met Ala Trp Phe Arg Gin Thr Pro Gly Lys Glu Arg Asp lie Val 35 40 45

Ala Ala He Ser Ser Ser Leu Gly Arg Thr Tyr Tyr Ala Asp Ser Val 50 55 60

Lys Gly Arg Phe Gin lie Leu Arg Asp Asn Ala Lys Glu Thr Val Trp 65 70 75 80

Leu Gin Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Val Tyr He Cys 85 90 95

Ala Ala Ser Arg Ser Leu Asn Leu Ala Tyr Thr Thr Lys Pro Tyr Asp 100 105 110

Tyr Trp Gly Gin Gly Thr Gin Val Thr Val Ser Ser 115 120 <210> 82 <211> 133

<212> PRT <213> Lama glama <400> 82

Glu Val Gin Leu Val Glu Ser Gly Gly Gly LeU Val Gin Ala Gly Gly 15 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Asp Phe Glu Asp Tyr 20 25 30

Asp Met Gly Trp Phe Arg Gin Ala Pro Gly Lys Glu Arg Glu Gly Val 35 40 45

Ser Tyr He Ser Ser Ser Asp Gly Ser Thr Tyr Tyr Thr Asp Ser Val 50 55 60

Lys Gly Arg Phe Thr He Ser Sen Asp Asn Ala Lys Asn Thr Val Tyr 65 70 75 80

Leu Gin Met Asn Ser Leu He Pro Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Ala Arg Pro Trp Thr Arg Arg Val Tyr Gly Ser Ser Trp Leu Ala 100 105 110

Arg Ser Leu Asp Glu Tyr Glu Tyr Asp Tyr Trp Gly Gin Gly Thr Gin 115 120 125

Val Thr Val Ser Ser 130 <210> 83 <211> 123

<212> PRT <213> Lama glama <400> 83

Glu Val Gin Leu Val Glu Ser Gly Gly Gly Leu Val Gin Val Gly Asp 15 10 15

Ser Leu Arg Leu Ser CyS Ala Ala Ser Gly His Thr Phe Ser Thr Tyr 20 25 30

Leu Met Val Gly Trp Phe Arg Gin Thr Pro Gly Lys Glu Arg Glu Phe 35 40 45

Ala Ala Val lie Trp Ser Ser Gly Asp Thr Ala Tyr Ala Asp Ser Ala 50 55 60

Lys Gly Arg Phe Thr He Ser Arg Asp Asn Ala Lys Asn Thr Val Tyr 65 70 75 80

Leu Gin Met Asn Ser Leu Lys Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Gly Ser Tyr Gly Gly Asn Tyr Tyr He Pro Gly Phe Tyr Glu Asp 100 105 110

Trp Gly Gin Gly Thr Gin Val Thr Val Ser Ser 115 120 <210> 84 <211> 126

<212> PRT <213> Lama glama <400> 84

Glu Val Gin Leu Met Glu Ser Gly Gly Gly Leu Val Gin Pro Gly Gly 1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Ala Leu Asp Tyr Tyr 20 25 30

Ala He Gly Trp Phe Arg Gin Val Pro Gly Lys Glu Arg Glu Gly Val 35 40 45

Ser Cys He Ser Ser Ser Asp Gly He Thr Tyr Tyr Val Asp Ser Val 50 55 60

Lys Gly Arg Phe Thr He Ser Arg Asp Asn Ala Lys Asn Thr Val Tyr 65 70 75 80

Leu Gin Met Asn Ser Leu Lys Pro Gly Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Thr Asp Ser Gly Gly Tyr He Asp Tyr Asp Cys Met Gly Leu Gly 100 105 110

Tyr Asp Tyr Trp Gly Gin Gly Thr Gin Val Thr Val Ser Ser 115 120 125 <210> 85 <211> 126

<212> PRT <213> Lama glama <400> 85

Glu Val Gin Leu Val Glu Ser Gly Gly Gly Leu Val Gin Pro Gly Gly 15 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Ala Leu Asp Tyr Tyr 20 25 30

Ala lie Gly Trp Phe Arg Gin Ala Pro Gly Lys Glu Arg Glu Gly Val 35 40 45

Ser Cys He Ser Ser Ser Asp Gly He Thr Tyr Tyr Val Asp Ser val 50 55 60

Lys Gly Arg Phe Thr He Ser Arg Asp Asn Ala Lys Asn Thr Val Tyr 65 70 75 80

Leu Gin Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Thr Asp Ser Gly Gly Tyr He Asp Tyr Asp Cys Met Gly Leu Gly 100 105 110

Tyr Asp Tyr Trp Gly Gin Gly Thr Gin Val Thr Val Ser Ser 115 120 125 <210> 86 <211> 123

<212> PRT <213> Lama glama <400> 86

Glu Val Gin Leu Val Glu Ser Gly Gly Gly Leu Val Gin Val Gly Asp 15 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Arg Thr Phe Ser Thr Tyr 20 25 30

Leu Met Val Gly Trp Phe Arg Gin Ala Pro Gly Lys Glu Arg Glu Phe 35 40 45

Ala Ala Val He Trp Ser Ser Gly Asp Thr Ala Tyr Ala Asp Ser Val 50 55 60

Lys Gly Arg Phe Thr He Ser Arg Asp Asn Ala Lys Asn Thr Val Tyr 65 70 75 80

Leu Gin Met Asn Ser Leu Lys Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Glv Ser Tvr Asn Glv Asn Tvr Tvr He Pro Glv Phe Tvr Lvs Asn

100 105 ITO

Trp Gly Gin Gly Thr Gin Val Thr Val Ser Ser 115 120 <210> 87 <211> 129

<212> PRT <213> Artificial <220> <223> Mutated Lama sequence <400> 87

Glu Val Gin Leu Val Glu Ser Gly Gly Gly Leu Val Gin Ala Gly Gly 15 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Leu Asp Asp Tyr 20 25 30

Ala lie Gly Trp Phe Arg Gin Ala Pro Gly Lys Glu Arg Glu Gly Val 35 40 45

Ser Ser lie Arg Asp Asn Asp Gly Ser Thr Tyr Tyr Ala Asp Ser Val 50 55 60

Lys Gly Arg Ser Thr lie Ser Ser Asp Asn Asp Lys Asn Thr Val Tyr 65 70 75 80

Leu Gin Met Asp Ser Leu Lys Pro Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Ala Val Pro Ala Gly Arg Leu Arg Tyr Gly Glu Gin Trp Tyr Pro 100 105 110

He Tyr Glu Tyr Asp Ala Trp Gly Gin Gly Thr Leu Val Thr Val Ser 115 120 125

Ser <210> 88 <211> 129

<212> PRT <213> Artificial <220> <223> Mutated Lama sequence <400> 88

Glu Val Gin Leu Val Glu Ser Gly Gly Gly Leu Val Gin Ala Gly Gly 1 5 10 15 τ λ,, λ τ Λ,, r*,·,,·, a l — ,. τΐ χ ipu»» t λ ocl ueii .nxij ueu oei t-ya Λ±α vax oex oxy xxts xxxx xieu A£>_p xyx 20 25 30

Ala Val Gly Trp Phe Arg Gin Ala Pro Gly Lys Glu Arg Glu Gly Val 35 40 45

Ser Thr He Arg Asp Asn Asp Gly Ser Thr Tyr Tyr Ala Asp Ser Val 50 55 60

Lys Gly Arg Phe Thr He Ser Ser ASp Asn Asp Lys Asn Thr Val Tyr 65 70 75 80

Leu Gin Met Asn Ser Leu Lys Pro Lys Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Ala Val Pro Ala Gly Arg Leu Arg Phe Gly Glu Gin Trp Tyr Pro 100 105 110

He Tyr Glu Tyr Asp Ala Trp Gly Gin Gly Thr Leu Val Thr Val Ser 115 120 125

Ser <210> 89 <211> 129

<212> PRT <213> Artificial <220> <223> Mutated Lama sequence <400> 89

Glu Val Gin Leu Val Glu Ser Gly Gly Gly Gin Ala Gin Ala Gly Gly 15 10 15

Ser Leu Arg Leu Ser Cys Ala Val Ser Gly Phe Thr Leu Asp Asp Tyr 20 25 30

Ala He Gly Trp Phe Arg Gin Ala Pro Gly Lys Glu Arg Glu Gly Val 35 40 45

Ser Ser He Arg Asp Asn Asp Gly Ser Thr Tyr Tyr Thr Asp Ser val 50 55 60

Lys Gly Arg Phe Thr He Ser Ser Asp Asn Asp Lys Asn Thr Val Tyr 65 70 75 80

Leu Gin Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Ala Val Pro Ala Gly Arg Leu Arg Phe Gly Glu Gin Trp Tyr Pro 100 105 110 lie Tyr Glu Tyr Asp Ala Trp Gly Gin Gly Thr Leu Val Thr Val Ser 115 120 125

Ser <210> 90 <211> 129

<212> PRT <213> Artificial <220> <223> Mutated Lama sequence <400> 90

Glu Val Gin Leu Val Glu Ser Gly Gly Gly Leu Val Gin Ala Gly Gly 15 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly lie Thr Leu Asp Asp Tyr 20 25 30

Ala He Gly Trp Phe Arg Gin Ala Pro Gly Lys Glu Arg Val Gly Val 35 40 45

Ser Ser He Arg Asp Asn Asp Gly Ser Thr Tyr Tyr Ala Asp Ser Val 50 55 60

Lys Gly Arg Phe Thr He Ser Ser Asp Asn Asp Lys Asn Thr Val Tyr 65 70 75 80

Leu Gin Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Ala Val Pro Ala Gly Arg Leu Arg Phe Gly Glu Gin Trp Tyr Pro 100 105 110

He Tyr Glu Tyr Asp Ala Trp Gly Gin Gly Thr Leu Val Thr Val Ser 115 120 125

Ser <210> 91 <211> 129

<212> PRT <213> Artificial <220> <223> Mutated Lama sequence <400> 91

Glu Val Gin Leu Val Glu Ser Gly Gly Gly Leu Val Gin Ala Gly Gly 1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Leu Thr Leu Asp Asp Tyr 20 25 30

Ala He Gly Trp Phe Arg Gin Ala Pro Gly Lys Glu Arg Glu Gly Val 35 40 45

Ser Ser lie Arg Asp Asn Asp Gly Ser Thr Tyr Tyr Ala Asp Ser Val 50 55 60

Lys Gly Arg Phe Thr He Ser Ser Asp Asn Asp Lys Asn Thr Val Tyr 65 70 75 80

Leu Gin Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Ala Val Pro Ala Gly Arg Leu Arg Phe Gly Glu Gin Trp Tyr Pro 100 105 110

He Tyr Glu Tyr Asp Ala Trp Gly Gin Gly Thr Leu val Thr Val Ser 115 120 125

Ser <210> 92 <211> 129

<212> PRT <213> Artificial <220> <223> Mutated Lama sequence <400> 92

Gin Val Gin Leu Val Glu Ser Gly Gly Gly Leu Val Gin Ala Gly Gly 15 10 15

Ser Leu Arg Leu Ser Cys Ala Val Ser Gly Phe Thr Leu Asp Asp Tyr 20 25 30

Ala He Gly Trp Phe Arg Gin Ala Pro Gly Lys Glu Arg Glu Gly Val 35 40 45

Ser Ser He Arg Asp Asn Asp Gly Ser Thr Tyr Tyr Ala Asp Ser Val 50 55 60

Lys Gly Arg Phe Thr lie Ser Ser Asp Asn Asp Gin Asn Thr Val Tyr 65 70 75 80

Leu Gin Met Asn Ser Leu Lys Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Ala Val Pro Ala Gly Arg Leu Arg Phe Gly Glu Gin Trp Tyr Pro 100 105 110

He Tyr Glu Tyr Asp Ala Trp Gly Gin Gly Thr Leu Val Thr Val Ser 115 120 125

Ser <210> 93 <211> 129

<212> PRT <213> Artificial <220> <223> Mutated Lama sequence <400> 93

Glu Val Gin Leu Val Glu Ser Gly Gly Gly Leu Val Gin Ala Gly Gly 15 10 15

Ser Leu Arg Leu Ser Cys Ala lie Ser Gly Phe Thr Leu Val Asp Tyr 20 25 30

Ala He Gly Trp Phe Arg Gin Ala Pro Gly Lys Glu Arg Glu Gly Val 35 40 45

Ser Ser He Arg Asp Asn Asp Gly Ser Thr Tyr Tyr Ala Asp Ser Val 50 55 60

Lys Gly Arg Phe He lie Ser Ser Asp Asn Asp Lys Asn Thr Val Tyr 65 70 75 80

Leu Gin Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Ala Val Pro Ala Gly Arg Leu Arg Phe Gly Glu Gin Trp Tyr Pro 100 105 110

He Tyr Glu Tyr Asp Ala Trp Gly Gin Gly Thr Leu Val Thr Val Ser 115 120 125

Sen <210> 94 <211> 129

<212> PRT <213> Artificial <220> <223> Mutated Lama sequence <400> 94

Glu Val Gin Leu Met Glu Ser Gly Gly Gly Leu Val Gin Ala Gly Gly 1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly He Thr Leu Asp Asp Tyr 20 25 30

Ala He Gly Trp Phe Arg Gin Ala Pro Gly Lys Glu Arg Glu Gly Val 35 40 45

Ser Ser He Arg Asp Asn Asp Gly Ser Thr Tyr Tyr Ala Asp Ser Val 50 55 60

Lys Gly Arg Phe Thr lie Ser Ser Asp Asn Asp Lys Asn Thr Val Asn 65 70 75 80

Leu Gin Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Ala Val Pro Ala Gly Arg Leu Arg Phe Gly Glu Gin Trp Tyr Pro 100 105 110

He Tyr Glu Tyr Asp Ala Trp Gly Gin Gly Thr Leu val Thr Val Ser 115 120 125

Ser <210> 95 <211> 129

<212> PRT <213> Artificial <220> <223> Mutated Lama sequence <400> 95

Glu Val Gin LeU Val Glu Ser Gly Gly Gly Leu Val Gin Ala Gly Gly 15 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Leu Asp Asp Tyr 20 25 30

Ala He Gly Trp Phe Arg Gin Ala Pro Gly Lys Glu Arg Glu Gly Val 35 40 45

Ser Ser He Arg Asp Asn Asp Gly Ser Thr Tyr Tyr Ala Asp Ser Val 50 55 60

Lys Gly Arg Phe Thr He Ser Ser Asp Asn Asp Lys Asn Thr Val Tyr 65 70 75 80

Leu Gin Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Ala Val Pro Ala Gly Arg Leu Arg Tyr Gly Glu Gin Trp Tyr Pro 100 105 110 lie Tyr Glu Tyr Asp Ala Trp Gly Gin Gly Thr Leu Val Thr Val Ser 115 120 125

Ser <210> 96 <211> 129

<212> PRT <213> Artificial <220> <223> Mutated Lama sequence <400> 96

Glu Val Gin LeU Val Glu Ser Gly Gly Gly Leu Val Gin Ala Gly Gly 15 10 15

Ser Leu Arg Leu Ser Cys Ala Val Ser Gly Phe Thr Leu Asp Asp Tyr 20 25 30

Ala lie Gly Trp Phe Arg Gin Ala Pro Gly Lys Glu Arg Glu Gly Val 35 40 45

Ser Ser He Arg Glu Asn Asp Gly Ser Thr Tyr Tyr Ala Asp Ser Val 50 55 60

Gin Gly Arg Phe Thr He Ser Ser Asp Asn Asp Lys Asn Thr Val Tyr 65 70 75 80

Leu Arg Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Ala Val Pro Ala Gly Arg Leu Arg Phe Gly Glu Gin Trp Tyr Pro 100 105 110

He Tyr Glu Tyr Asp Ala Trp Gly Gin Gly Thr Leu Val Thr Val Ser 115 120 125

Ser <210> 97 <211> 129

<212> PRT <213> Artificial <220> <223> Mutated Lama sequence <400> 97

Glu val Gin Met Val Glu Ser Gly Gly Gly Leu Val Gin Ala Gly Gly 15 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Leu Thr Leu Asp Asp Tyr 20 25 30

Ala He Gly Trp Phe Arg Gin Ala Pro Gly Lys Glu Arg Glu Gly Val 35 40 45

Ser Ser lie Arg Asp Asn Asp Gly Ser Tht Tyr Tyr Ala Asp Ser Val 50 55 60

Lys Gly Arg Phe Thr lie Ser Ser Asp Asn Asp Lys Asn Thr Val Tyr 65 70 75 80

Leu Gin Met Asn Gly Leu Lys Pro Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Ala Val Pro Ala Gly Arg Leu Arg Phe Gly Glu Gin Trp Tyr Pro 100 105 110 lie Tyr Glu Tyr Asp Ala Trp Gly Gin Gly Thr Leu Val Thr Val Ser 115 120 125

Ser <210> 98 <211> 129

<212> PRT <213> Artificial <220> <223> Mutated Lama sequence <400> 98

Glu Val Gin Leu Val Glu Ser Gly Gly Gly Leu Val Gin Ser Gly Gly 15 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Leu Thr Leu Asp Asp Tyr 20 25 30

Ala He Gly Trp Phe Arg Gin Ala Pro Gly Lys Glu Arg Glu Gly Val 35 40 45

Ser Ser He Arg Asp Asn Asp Gly Ser Thr Tyr Tyr Ala Asp Ser Val 50 55 60

Lys Gly Arg Phe Thr He Ser Ser Asp Asn Asp Lys Asn Thr Val Tyr 65 70 75 80

Leu Gin Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Ala Val Pro Ala Gly Arg Leu Arg Phe Gly Glu Gin Trp Tyr Pro 100 105 110 lie Tyr Glu Tyr Asp Ala Trp Gly Gin Gly Thr Leu Val Thr Val Ser 115 120 125

Ser <210> 99 <211> 129

<212> PRT <213> Artificial <220> <223> Mutated Lama sequence <400> 99

Glu Val Gin Leu Val Glu Ser Gly Gly Gly Leu Val Gin Ala Gly Gly 15 10 15

Ser Leu Arg Leu Ser Cys Ala Val Ser Gly Phe Thr Leu Asp Asp Tyr 20 25 30

Ala lie Gly Trp Phe Arg Gin Ala Pro Gly Lys Glu Arg Glu Gly Val 35 40 45

Ser Ser lie Arg Asp Asn Asp Gly Ser Thr Tyr Tyr Ala Asp Ser Val 50 55 60

Lys Gly Arg Ser Thr lie Ser Ser Asp Asn Asp Lys Asn Thr Val Tyr 65 70 75 80

Leu Gin Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Ala Val Pro Ala Gly Arg Leu Arg Phe Gly Glu Qin Trp Tyr Pro 100 105 110

He Tyr Glu Tyr Asp Ala Trp Gly Gin Gly Thr Leu Val Thr Val Ser 115 120 125

Ser <210> 100 <211> 129

<212> PRT <213> Artificial <220> <223> Mutated Lama sequence <400> 100

Glu Val Gin Leu Val Glu Ser Gly Gly Gly Leu Val Gin Ala Gly Gly 15 10 15

Ser Leu Arg Leu Ser Cys Ala Val Ser Gly He Thr Leu Asp Asp Tyr 20 25 30

Ala He Gly Trp Phe Arg Gin Ala Pro Gly Lys Glu Arg Glu Gly Val 35 40 45

Ser Ser He Arg Asp Asn Asp Gly Ser Thr Tyt Tyr Ala Asp Ser Val 50 55 60

Lys Trp Arg Phe Thr lie Ser Ser ASp Asn Asp Lys Asn Thr Val Tyr 65 70 75 80

Leu Gin Met Asn Ser Met Lys Pro Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Axa Axa vax pro Axa uxy Arg xeu Arg Fne uxy uxu uxn irp ryr Fro 100 105 110 lie Tyr Glu Tyr Asp Ala Trp Gly Gin Gly He Leu Val Thr Val Ser 115 120 125

Ser <210> 101 <211> 129

<212> PRT <213> Artificial <220> <223> Mutated Lama sequence <400> 101

Glu Val Gin Leu Val Glu Ser Gly Gly Gly Leu Val Gin Ala Gly Gly 15 10 15

Phe Leu Arg Leu Thr Cys Ala Val Ser Gly Phe Thr Leu Asp Asp Tyr 20 25 30

Ala He Gly Trp Phe Arg Gin Ala Pro Gly Lys Glu Arg Glu Gly Val 35 40 45

Ser Ser He Arg Asp Asn Asp Gly Ser Thr Tyr Tyr Ala Asp Ser Val 50 55 60

Lys Gly Arg Phe Thr He Ser Ser ASp Asn Val Lys Asn Thr Val Tyr 65 70 75 80

Leu Gin Met Asn Ser Leu Lys Pro Glu ASp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Ala Val Pro Ala Gly Arg Leu Arg Phe Gly Glu Gin Trp Tyr Pro 100 105 110

He Tyr Glu Tyr Asp Ala Trp Gly Gin Gly Thr Leu Val Thr Val Ser 115 120 125

Ser <210> 102 <211> 129

<212> PRT <213> Artificial <220> <223> Mutated Lama sequence <400> 102

Glu Val Gin Leu Leu Glu Ser Gly Gly Gly Leu Val Gin Ala Gly Gly 1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Leu Asp Asp Tyr 20 25 30

Ala lie Gly Trp Phe Arg Gin Ala Pro Gly Lys Glu Arg Glu Gly Val 35 40 45

Ser Ser He Arg Asp Asn Asp Gly Ser Thr Tyr Tyr Ala Asp Ser Val 50 55 60

Lys Gly Arg Phe Thr He Ser Ser Asp Asn Asp Lys Asn Thr Val Tyr 65 70 75 80

Leu Gin Met Asn Ser Leu Lys Pro Glu ASp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Ala Val Pro Ala Gly Arg Leu Arg He Gly Glu Gin Trp Tyr Pro 100 105 110

He Tyr Glu Tyr Asp Ala Trp Gly Gin Gly Thr Leu Val Thr Val Ser 115 120 125

Ser <210> 103 <211> 129

<212> PRT <213> Artificial <220> <223> Mutated Lama sequence <400> 103

Glu Val His Leu Val Glu Ser Gly Gly Gly Leu Val Gin Ala Gly Gly 15 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Leu Thr Leu Asp Asp Tyr 20 25 30

Ala He Gly Trp Phe Arg Gin Ala Pro Gly Val Glu Arg Glu Gly Val 35 40 45

Ser Ser He Arg Asp Asn Asp Gly Ser Thr Tyr Tyr Ala Asp Ser Val 50 55 60

Lys Gly Arg Phe Thr He Ser Ser Asp Asn Asp Lys Asn Thr Val Tyr 65 70 75 80

Leu Gin Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Ala Val Pro Ala Gly Arg Leu Arg Phe Gly G1U Gin Trp Tyr Pro 100 105 110 lie Tyr Glu Tyr Asp Ala Trp Gly Gin Gly Thr Leu Val Thr Val Ser 115 120 125

Ser <21 0> 104 <211> 129

<212> PRT <213> Artificial <220> <223> Mutated Lama sequence <400> 104

Glu Val Gin Leu Val Glu Ser Gly Gly Gly Leu Val Gin Ala Gly Gly 1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Leu Asp Asp Tyr 20 25 30

Ala lie Gly Trp Phe Arg Gin Ala Pro Gly Lys Asp Arg Glu Gly Val 35 40 45

Ser Ser He Arg Asp Asn Asp Gly Ser Thr Tyr Tyr Ala Asp Ser Val 50 55 60

Lys Gly Arg Phe Thr lie Ser Ser Asp Asn Asp Lys Asn Thr Val Tyr 65 70 75 80

Leu Gin Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Ala Val Pro Ala Gly Arg Leu Arg Tyr Gly Glu Gin Trp Tyr Pro 100 105 110

He Tyr Glu Tyr Asp Ala Trp Gly Gin Gly Thr Val Val Thr Val Ser 115 120 125

Ser <210> 105 <211> 129

<212> PRT <213> Artificial <220> <223> Mutated Lama sequence <400> 105

Glu Val Gin Leu val Glu Ser Gly Gly Gly Leu Val Gin Ala Gly Gly 15 10 15

Thr Leu Arg LeU Ser Cys Ala Val Ser Gly Phe Thr Leu Asp Asp Tyr 20 25 30

Ala He Gly Trp Phe Arg Gin Ala Pro Gly Lys Glu Arg Glu Gly Val 35 40 45

Ser Ser He Arg Asp Ash Asp Gly Ser Thr Tyr Tyr Ala Asp Ser Val 50 55 60

Lys Gly Arg Phe Thr lie Ser Ser Asp Ash Asp Lys Asn Thr val Tyr 65 70 75 80

Leu Gin Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Ala Val Pro Ala Gly Arg Leu Arg Phe Gly Glu Gin Trp Tyr Pro 100 105 110

He Tyr Glu Tyr Asp Ala Trp Gly Gin Gly lie Leu Val Thr Val Ser 115 120 125

Ser <210> 106 <211> 129

<212> PRT <213> Artificial <220> <223> Mutated Lama sequence <400> 106

Glu Val Gin Leu Val Glu Ser Gly Gly Gly Leu Val Gin Ala Gly Gly 15 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Leu Asp Asp Tyr 20 25 30

Ala He Gly Trp Phe Arg Gin Ala Pro Gly Lys Glu Arg Glu Gly Val 35 40 45

Ser Ser He Arg Asp Asn Asp Gly Ser Thr Tyr Tyr Ala Asp Ser Val 50 55 60

Lys Gly Arg Phe He He Ser Ser Asp Asn Asp Lys Asn Thr Val Tyr 65 70 75 80

Leu Gin Met Asn Asn Leu Lys Pro Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Ala Val Pro Ala Gly Arg Leu Arg Tyr Gly Glu Gin Trp Tyr Pro 100 105 110 lie Tyr Glu Tyr Asp Ala Trp Gly Gin Gly Thr Leu Val Thr Val Ser 115 120 125

Ser <210> 107 <211> 129

<212> PRT <213> Artificial <220> <223> Mutated Lama sequence <400> 107

Glu Val Gin Leu Val Glu Ser Gly Gly Gly Leu Val Gin Ala Gly Gly 15 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Leu Asp Asp Tyr 20 25 30

Ala lie Gly Trp Phe Arg Gin Ala Pro Gly Lys Glu Arg Glu Gly Val 35 40 45

Ser Ser He Arg Asp Asn Asp Gly Ser Thr Tyr Tyr Ala Asp Ser Val 50 55 60

Lys Gly Arg Ser Thr He Ser Ser Asp Asn Ala Arg Asn Thr Val Phe 65 70 75 80

Leu Gin Met ASrt Ser Leu LyS Pro Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Ala Val Pro Ala Gly Arg Leu Arg Phe Gly Glu Gin Trp Tyr Pro 100 105 110

He Tyr Glu Tyr Asp Ala Trp Gly Gin Gly Thr Leu Val Thr Val Ser 115 120 125

Ser <210> 108 <211> 129

<212> PRT <213> Artificial <220> <223> Mutated Lama sequence <400> 108

Glu Glu Leu Leu Val Glu Ser Gly Gly Gly Ser Val Gin Ala Gly Gly 15 10 15

Ser Leu Lys Leu Ser Cys Ala Ala Ser Gly Leu Thr Leu Asp Asp Tyr 20 25 30

Ala Tie Glv Trn Phe Am Gin Ala Pro Glv Lvs Gin Am Glu Glv Val 35J ^40 J 45’

Ser Ser lie Arg Asp Asn Asp Gly Ser Thr Tyr Tyr Ala Asp Ser Val 50 55 60

Lys Gly Arg Phe Thr He Ser Ser Asp Asn Asp Lys Asn Thr Val Tyr 65 70 75 80

Leu Gin Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Ala Val Pro Ala Gly Arg Leu Arg Phe Gly Glu Gin Trp Tyr Pro 100 105 110

He Tyr Glu Tyr Asp Ala Trp Gly Gin Gly Thr Leu Val Thr Val Ser 115 120 125

Ser <210> 109 <211> 129

<212> PRT <213> Artificial <220> <223> Mutated Lama sequence <400> 109

Glu Val Gin Leu Val Glu Ser Gly Gly Gly Leu Val Gin Ala Gly Gly 15 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Leu Thr Leu Asp Asp Tyr 20 25 30

Ala He Gly Trp Phe Arg Gin Ala Pro Gly Lys Glu Arg Glu Gly Val 35 40 45

Ser Ser He Arg Asp Asn Asp Gly Ser Arg Tyr Tyr Ala Asp Ser Val 50 55 60

Lys Gly Arg Phe Thr He Ser Ser Asp Asn Asp Lys Asn Thr Val Tyr 65 70 75 80

Leu Gin Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Ala Val Pro Ala Gly Arg Leu Arg Phe Gly Glu Gin Trp Tyr Pro 100 105 110

He Tyr Glu Tyr Asp Ala Trp Gly Gin Gly Thr Leu Val Thr Val Ser 115 120 125

Ser <210> 110 <211> 129

<212> PRT <213> Artificial <220> <223> Mutated Lama sequence <400> 110

Glu Val Gin LeU Val Glu Ser Gly Gly Ser Leu Val Gin Ala Gly Gly 15 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Leu Thr Leu Asp Asp Tyr 20 25 30

Ala lie Gly Trp Phe Arg Gin Ala Pro Gly Lys Glu Arg Glu Gly Val 35 40 45

Ser Ser He Arg Asp Asn Asp Gly Ser Thr Tyr Tyr Ala Asp Ser Val 50 55 60

Lys Gly Arg Phe Thr He Ser Ser Asp Asn Asp Lys Asn Thr Val Tyr 65 70 75 80

Leu Gin Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Ala Val Pro Ala Gly Arg Leu Arg Phe Gly Glu Gin Trp Tyr Pro 100 105 110 lie Tyr Glu Tyr Asp Ala Trp Gly Gin Gly Thr Leu Val Thr Val Ser 115 120 125

Ser <210> 111 <211> 129

<212> PRT <213> Artificial <220> <223> Mutated Lama sequence <400> 111

Glu Val Gin Leu Val Glu Ser Gly Gly Ser Leu Val Gin Ala Gly Gly 15 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Leu Thr Leu Asp Asp Tyr 20 25 30

Ala He Gly Trp Phe Arg Gin Ala Pro Gly Lys Glu Arg Glu Gly Val 35 40 45

Ser Ser lie Arg Asp Asn Asp Gly Ser Thr Tyr Tyr Ala Asp Ser Val 50 55 60

Lys Gly Arg Phe Thr He Ser Ser Asp Asn Asp Lys Asn Thr Val Tyr 65 70 75 80

Leu Glh Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Ala Val Pro Ala Gly Arg Leu Arg Phe Gly Glu Gin Trp Tyr Pro 100 105 110

He Tyr Glu Tyr Asp Ala Trp Gly Gin Gly Thr Leu Val Thr Val Ser 115 120 125

Ser <210> 112 <211> 129

<212> PRT <213> Artificial <220> <223> Mutated Lama sequence <400> 112

Glu Val Gin Leu Val Glu Ser Gly Gly Gly Leu Val Gin Pro Gly Gly 1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Leu Asp Asp Tyr 20 25 30

Ala He Gly Trp Phe Arg Glu Ala Pro Gly Lys Glu Arg Glu Gly Val 35 40 45

Ser Ser He Arg Asp Asn Gly Gly Ser Thr Tyr Tyr Ala Asp Ser Val 50 55 60

Lys Gly Arg Phe Thr He Ser Ser Asp Asn Ser Lys Asn Thr Val Tyr 65 70 75 80

Leu Gin Met Asn Ser Leu Arg Pro Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Ala Val Pro Ala Gly Arg Leu Arg Phe Gly Glu Gin Trp Tyr Pro 100 105 110 lie Tyr Glu Tyr Asp Ala Trp Gly Gin Gly Thr Leu Val Thr Val Ser 115 120 125

Ser <210> 113 <211> 129

<212> PRT <213> Artificial <220> <223> Mutated Lama sequence <400> 113

Glu Val Gin Leu Val Glu Ser Gly Gly Gly Leu Val Gin Pro Gly Gly 15 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Leu Asp Asp Tyr 20 25 30

Ala lie Gly Trp Phe Arg Gin Ala Pro Gly Lys Glu Arg Glu Gly Val 35 40 45

Ser Ser He Arg Asp Asn Gly Gly Ser Thr Tyr Tyr Ala Asp Ser Val 50 55 60

Lys Gly Arg Phe Thr He Ser Ser Asp Asn Ser Lys Asn Thr Val Tyr 65 70 75 80

Leu Gin Met Asn Ser Leu Arg Pro Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Ala Val Pro Ala Gly Arg Leu Arg Tyr Gly Glu Gin Trp Tyr Pro 100 105 110

He Tyr Glu Tyr Asp Ala Trp Gly Gin Gly Thr Leu Val Thr Val Ser 115 120 125

Ser <210> 114 <211> 129

<212> PRT <213> Artificial <220> <223> Mutated Lama sequence <400> 114

Glu Val Gin Leu val Glu Ser Gly Gly Gly Leu Val Gin Pro Gly Gly 15 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr He Asp Asp Tyr 20 25 30

Ala He Gly Trp Phe Arg Gin Ala Pro Gly Lys Glu Arg Glu Gly Val 35 40 45

Ser Ser He Arg Asp Asn Gly Gly Ser Thr Tyr Tyr Ala Asp Ser Val 50 55 60

Lys Gly Arg Phe Thr lie Ser Ser Asp Asn Ser Lys Asn Thr val Tyr 65 70 75 80

Leu Gin Met Asn Ser Leu Arg pro Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Ala Val Pro Ala Gly Arg Leu Arg Phe Gly Glu Gin Trp Tyr Pro 100 105 110 lie Tyr Glu Tyr Asp Ala Trp Gly Gin Gly Thr Leu Val Thr Val Ser 115 120 125

Ser <210> 115 <211> 129

<212> PRT <213> Artificial <220> <223> Mutated Lama sequence <400> 115

Glu Val Gin LeU Val Glu Ser Gly Gly Gly Leu Val Gin Pro Gly Gly 15 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr He Asp Asp Tyr 20 25 30

Ala He Gly Trp Phe Arg Gin Ala Pro Gly Lys Glu Arg Glu Gly Val 35 40 45

Ser Ser He Arg Asp Asn Gly Gly Ser Thr Tyr Tyr Ala Asp Ser Val 50 55 60

Lys Gly Arg Phe Thr He Ser Ser Asp Asn Ser Lys Asn Thr Val Tyr 65 70 75 80

Leu Gin Met Asn Ser Leu Arg Pro Glu ASp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Ala Val Pro Ala Gly Arg Leu Arg Tyr Gly Glu Gin Trp Tyr Pro 100 105 110 lie Tyr Glu Tyr Asp Ala Trp Gly Gin Gly Thr Leu Val Thr Val Ser 115 120 125

Ser <210> 116 <211> 129

<212> PRT <213> Artificial <220> <223> Mutated Lama sequence <400> 116

Glu Val Gin Leu Val Glu Ser Gly Gly Gly Leu Val Gin Pro Gly Gly 15 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly lie Thr Leu Asp Asp Tyr 20 25 30

Ala He Gly Trp Phe Arg Gin Ala Pro Gly Lys Glu Arg Glu Gly Val 35 40 45

Ser Ser He Arg Asp Asn Gly Gly Ser Thr Tyr Tyr Ala Asp Ser Val 50 55 60

Lys Gly Arg Phe Thr He Ser Ser Asp Asn Ser Lys Asn Thr Val Tyr 65 70 75 80

Leu Gin Met Asn Ser Leu Arg Pro Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Ala Val Pro Ala Gly Arg Leu Arg Phe Gly Glu Gin Trp Tyr Pro 100 105 110 lie Tyr Glu Tyr Asp Ala Trp Gly Gin Gly Thr Leu Val Thr Val Ser 115 120 125

Ser <210> 117 <211> 129

<212> PRT <213> Artificial <220> <223> Mutated Lama sequence <400> 117

Glu Val Gin Leu Val Glu Ser Gly Gly Gly Leu Val Gin Pro Gly Gly 15 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly He Thr Leu Asp Asp Tyr 20 25 30

Ala lie Gly Trp Phe Arg Gin Ala Pro Gly Lys Glu Arg Glu Gly Val 35 40 45

Ser Ser lie Arg Asp Asn Gly Gly Ser Thr Tyr Tyr Ala Asp Ser Val 50 55 60

Lys Gly Arg Phe Thr lie Ser Ser Asp Asn Ser Lys Asn Thr Val Tyr 65 70 75 80 T.en Qin Mai- Δβη Sar T.an Dra Qin .len Thf Ala Val Tur Tvr Γ*ντβ 85 * 9(Λ 95

Ala Ala Val Pro Ala Gly Arg Leu Arg Tyr Gly Glu Gin Trp Tyr Pro 100 105 110 lie Tyr Glu Tyr Asp Ala Trp Gly Gin Gly Thr LeU Val Thr Val Ser 115 120 125

Ser <210> 118 <211> 129

<212> PRT <213> Artificial <220> <223> Mutated Lama sequence <400> 118

Glu Val Gin Leu Val Glu Ser Gly Gly Gly Leu Val Gin Pro Gly Gly 15 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly lie Thr He Asp Asp Tyr 20 25 30

Ala He Gly Trp Phe Arg Gin Ala Pro Gly Lys Glu Arg Glu Gly Val 35 40 45

Ser Ser lie Arg Asp Asn Gly Gly Ser Thr Tyr Tyr Ala Asp Ser Val 50 55 60

Lys Gly Arg Phe Thr He Ser Ser Asp Asn Ser Lys Asn Thr Val Tyr 65 70 75 80

Leu Gin Met Asn Ser Leu Arg Pro Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Ala Val Pro Ala Gly Arg Leu Arg Phe Gly Glu Gin Trp Tyr Pro 100 105 110

He Tyr Glu Tyr Asp Ala Trp Gly Gin Gly Thr Leu Val Thr Val Ser 115 120 125

Ser <210> 119 <211> 129

<212> PRT <213> Artificial <220> <223> Mutated Lama sequence <400> 119

Glu Val Gin Leu Val Glu Ser Gly Gly Gly Leu Val Gin Pro Gly Gly 1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly lie Thr He Asp Asp Tyr 20 25 30

Ala He Gly Trp Phe Arg Gin Ala Pro Gly Lys G1U Arg Glu Gly Val 35 40 45

Ser Ser He Arg Asp Asn Gly Gly Ser Thr Tyr Tyr Ala Asp Ser Val 50 55 60

Lys Gly Arg Phe Thr He Ser Ser Asp ASn Ser Lys Asn Thr Val Tyr 65 70 75 80

Leu Glh Met ASn Ser Leu Arg Pro Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Ala Val Pro Ala Gly Arg Leu Arg Tyr Gly Glu Gin Trp Tyr Pro 100 105 110

He Tyr Glu Tyr Asp Ala Trp Gly Gin Gly Thr Leu Val Thr Val Ser 115 120 125

Ser <210> 120 <211> 129

<212> PRT <213> Artificial <220> <223> Mutated Lama sequence <400> 120

Glu val Gin Leu Val Glu Ser Gly Gly Gly Leu Val Gin Pro Gly Gly 15 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Leu Thr Leu Asp Asp Tyr 20 25 30

Ala He Gly Trp Phe Arg Gin Ala Pro Gly Lys Glu Arg Glu Gly Val 35 40 45

Ser Ser He Arg Asp Asn Gly Gly Ser Tht Tyr Tyr Ala Asp Ser Val 50 55 60

Lys Gly Arg Phe Thr He Ser Ser Asp Asn Ser Lys Asn Thr Val Tyr 65 70 75 80

Leu Gin Met Asn Ser Leu Arg Pro Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Ala Val Pro Ala Glv Arrr Laii Arrr Pha Glw (Glu Gin Trn Twr Pro- 100 105 J 110 lie Tyr Glu Tyr Asp Ala Trp Gly Gin Gly Thr Leu Val Thr Val Ser 115 120 125

Ser <210> 121 <211> 129

<212> PRT <213> Artificial <220> <223> Mutated Lama sequence <400> 121

Glu Val Gin Leu Val Glu Ser Gly Gly Gly Leu Val Gin Pro Gly Gly 15 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Leu Thr Leu Asp Asp Tyr 20 25 30

Ala He Gly Trp Phe Arg Gin Ala Pro Gly Lys Glu Arg Glu Gly Val 35 40 45

Ser Ser He Arg Asp Asn Gly Gly Ser Thr Tyr Tyr Ala Asp Ser Val 50 55 60

Lys Gly Arg Phe Thr He Ser Ser Asp Asn Ser Lys ASn Thr Val Tyr 65 70 75 80

Leu Gin Met Asn Ser Leu Arg Pro Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Ala Val Pro Ala Gly Arg Leu Arg Tyr Gly Glu Gin Trp Tyr Pro 100 105 110

He Tyr Glu Tyr Asp Ala Trp Gly Gin Gly Thr Leu Val Thr Val Ser 115 120 125

Ser <210> 122 <211> 129

<212> PRT <213> Artificial <220> <223> Mutated Lama sequence <400> 122

Glu Val Gin Leu Val Glu Ser Gly Gly Gly Leu Val Gin Pro Gly Gly 15 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Leu Thr lie Asp Asp Tyr 20 25 30

Ala He Gly Trp Phe Arg Gin Ala Pro Gly Lys Glu Arg Glu Gly Val 35 40 45

Ser Ser He Arg Asp Asn Gly Gly Ser Thr Tyr Tyr Ala Asp Ser Val 50 55 60

Lys Gly Arg Phe Thr He Ser Ser Asp Asn Ser Lys Asn Thr Val Tyr 65 70 75 80

Leu Gin Met Asn Ser Leu Arg Pro Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Ala Val Pro Ala Gly Arg Leu Arg Phe Gly Glu Gin Trp Tyr Pro 100 105 110

He Tyr Glu Tyr Asp Ala Trp Gly Gin Gly Thr Leu Val Thr Val Ser 115 120 125

Ser <210> 123 <211> 129

<212> PRT <213> Artificial <220> <223> Mutated Lama sequence <400> 123

Glu Val Gin Leu Val Glu Ser Gly Gly Gly Leu Val Gin Pro Gly Gly 1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Leu Thr lie Asp Asp Tyr 20 25 30

Ala He Gly Trp Phe Arg Gin Ala Pro Gly Lys Glu Arg Glu Gly Val 35 40 45

Ser Ser He Arg Asp Asn Gly Gly Ser Thr Tyr Tyr Ala Asp Ser Val 50 55 60

Lys Gly Arg Phe Thr He Ser Ser Asp Asn Ser Lys Asn Thr Val Tyr 65 70 75 80

Leu Gin Met Asn Ser Leu Arg Pro Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Ala Val Pro Ala Gly Arg Leu Arg Tyr Gly Glu Gin Trp Tyr Pro 100 105 110

He Tyr Glu Tyr Asp Ala Trp Gly Gin Gly Thr Leu Val Thr Val Ser

Ί 1 C 1 Ο Λ 1 OC

XXJ X^.J

Ser <210> 124 <211> 124

<212> PRT <213> Artificial <220> <223> Mutated Lama sequence <400> 124

Glu Val Gin Leu Val Glu Ser Gly Gly Gly Leu Val Gin Pro Gly Gly 15 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Asp Asp Tyr 20 25 30

Ala Leu Gly Trp Phe Arg Gin Ala Ala Gly Lys Glu Arg Glu Gly Val 35 40 45

Ser Cys lie Arg Cys Ser Asp Gly Ser Thr Tyr Tyr Ala Asp Ser Val 50 55 60

Lys Gly Arg Phe Thr He Ser Arg Asp Asn Ser Lys Asn Thr Val Tyr 65 70 75 80

Leu Gin Met Asn Ser Leu Arg Pro Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Ala Ser lie Val Pro Arg Ser Lys Leu Glu Pro Tyr Glu Tyr Asp 100 105 HO

Ala Trp Gly Gin Gly Thr Leu Val Thr Val Ser Ser 115 120 <210> 125 <211> 124

<212> PRT <213> Artificial <220> <223> Mutated Lama sequence <400> 125

Glu Val Gin Leu Val Glu Ser Gly Gly Gly Leu Val Gin Pro Gly Gly 15 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Asp Asp Tyr 20 25 30

Ala Leu Gly Trp Phe Arg Gin Ala Pro Gly Lys Glu Arg Glu Gly Val 35 40 45

Ser Cys lie Arg Cys Ser Asp Gly Ser Thr Tyr Tyr Ala Asp Ser Val 50 55 60

Lys Gly Arg Phe Thr He Ser Arg Asp Asn Ser Lys Asn Thr Val Tyr 65 70 75 80

Leu Gin Met Asn Ser Leu Arg Pro Glu Asp Thr Ala val Tyr Tyr Cys 85 90 95

Ala Ala Ser He Val Pro Arg Ser Lys Leu Glu Pro Tyr Glu Tyr Asp 100 105 110

Ala Trp Gly Gin Gly Thr Leu Val Thr Val Ser Ser 115 120 <210> 126 <211> 124

<212> PRT <213> Artificial <220> <223> Mutated Lama sequence <400> 126

Glu Val Gin Leu Val Glu Ser Gly Gly Gly Leu Val Gin Pro Gly Gly 15 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Asp Asp Tyr 20 25 30

Ala Leu Gly Trp Phe Arg Gin Ala Ala Gly Lys Glu Arg Glu Gly Val 35 40 45

Ser Cys lie Arg Cys Ser Asp Gly Ser Thr Tyr Tyr Ala Asp Ser Val 50 55 60

Lys Gly Arg Phe Thr He Ser Ser Asp Asn Ser Lys Asn Thr Val Tyr 65 70 75 80

Leu Gin Met Asn Ser Leu Arg Pro Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Ala Ser He Val Pro Arg Ser Lys Leu Glu Pro Tyr Glu Tyr Asp 100 105 110

Ala Trp Gly Gin Gly Thr Leu Val Thr Val Ser Ser 115 120 <210> 127 <211> 124

<212> PRT <213> Artificial <220> <223> Mutated Lama sequence <400> 127

Glu Val Gin Leu Val Glu Ser Gly Gly Gly Leu Val Gin Pro Gly Gly 15 10 15

Ser LeU Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Asp Asp Tyr 20 25 30

Ala Leu Gly Trp Phe Arg Gin Ala Pro Gly Lys Glu Arg Glu Gly Val 35 40 45

Ser Cys lie Arg Cys Ser Asp Gly Ser Thr Tyr Tyr Ala Asp Ser Val 50 55 60

Lys Gly Arg Phe Thr He Ser Ser Asp Asn Ser Lys Asn Thr Val Tyr 65 70 75 80

Leu Gin Met Asn Ser Leu Arg Pro Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Ala Ser He Val Pro Arg Ser Lys Leu Glu Pro Tyr Glu Tyr Asp 100 105 110

Ala Trp Gly Gin Gly Thr Leu Val Thr Val Ser Ser 115 120 <210> 128 <211> 124

<212> PRT <213> Artificial <220> <223> Mutated Lama sequence <400> 128

Glu Val Gin Leu Val Glu Ser Gly Gly Gly Leu Val Gin Pro Gly Gly 15 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Asp Asp Tyr 20 25 30

Ala Leu Gly Trp Phe Arg Gin Ala Ala Gly Lys Glu Arg Glu Gly Val 35 40 45

Ser Cys lie Arg Cys Ser Gly Gly Ser Thr Tyr Tyr Ala Asp Ser Val 50 55 60

Lys Gly Arg Phe Thr He Ser Ser Asp Asn Ser Lys Asn Thr Val Tyr 65 70 75 80

Leu Gin Met Asn Ser Leu Arg Pro Glu Asp Thr Ala val Tyr Tyr Cys 85 90 95

Ala Ala Ser lie Val Pro Arg Ser Lys Leu Glu Pro Tyr Glu Tyr Asp 100 105 110

Ala Trp Gly Gin Gly Thr Leu Val Thr Val Ser Ser 115 120 <210> 129 <211> 124

<212> PRT <213> Artificial <220> <223> Mutated Lama sequence <400> 129

Glu Val Gin Leu Val Glu Ser Gly Gly Gly Leu Val Gin Pro Gly Gly 15 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Asp Asp Tyr 20 25 30

Ala Leu Gly Trp Phe Arg Gin Ala Pro Gly Lys Glu Arg Glu Gly VU1 35 40 45

Ser Cys He Arg Cys Ser Gly Gly Ser Thr Tyr Tyr Ala Asp Ser Val 50 55 60

Lys Gly Arg Phe Thr He Ser Ser Asp Asn Ser Lys Asn Thr Val Tyr 65 70 75 80

Leu Gin Met Asn Ser Leu Arg Pro Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Ala Ser He Val Pro Arg Ser Lys Leu Glu Pro Tyr Glu Tyr Asp 100 105 110

Ala Trp Gly Gin Gly Thr Leu Val Thr Val Ser Ser 115 120 <210> 130 <211> 124

<212> PRT <213> Artificial <220> <223> Mutated Lama sequence <400> 130

Asp Val Gin Leu Val Glu Ser Gly Gly Gly Leu Val Gin Pro Gly Gly 1 5 10 15

Ser Leu Anr Leu Ser Cvs Ala Ala Ser Glv Phe Thr Phe Asn Asn Tvr 20 J 25 30^

Ala Leu Gly Trp Phe Arg Gin Ala pro Gly Lys Glu Arg Glu Gly Val 35 40 45

Ser Cys lie Arg Cys Ser Gly Gly Ser Thr Tyr Tyr Ala Asp Ser Val 50 55 60

Lys Gly Arg Phe Thr He Ser Ser Asp Asn Ser Lys Asn Thr Val Tyr 65 70 75 80

Leu Gin Met Asn Ser Leu Arg Pro Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Ala Ser He Val Pro Arg Ser Lys Leu Glu Pro Tyr Glu Tyr Asp 100 105 110

Ala Trp Gly Gin Gly Thr Leu Val Thr Val Ser Ser 115 120 <210> 131 <211> 126

<212> PRT <213> Artificial <220> <223> Mutated Lama sequence <400> 131

Glu Val Gin Leu Val Glu Ser Gly Gly Gly Leu Val Gin Pro Gly Gly 15 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Ala Leu Asp Tyr Tyr 20 25 30

Ala He Gly Trp Phe Arg Gin Ala Pro Gly Lys Glu Arg Glu Gly Val 35 40 45

Ser Cys He Ser Ser Ser Asp Gly He Thr Tyr Tyr Val Asp Ser Val 50 55 60

Lys Gly Arg Phe Thr He Ser Arg Asp Asn Ser Lys Asn Thr Val Tyr 65 70 75 80

Leu Gin Met Asn Ser Leu Arg Pro Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Thr Asp Ser Gly Gly Tyr He Asp Tyr Asp Cys Met Gly Leu Gly 100 105 110

Tyr Asp Tyr Trp Gly Gin Gly Thr Leu Val Thr Val Ser Ser 115 120 125 <210> 132 <211> 126

<212> PRT <213> Artificial <220> <223> Mutated Lama sequence <400> 132

Glu Val Gin Leu Val Glu Ser Gly Gly Gly Leu Val Gin Pro Gly Gly 1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Ala Leu Asp Tyr Tyr 20 25 30

Ala lie Gly Trp Phe Arg Gin Ala Pro Gly Lys Glu Arg Glu Gly Val 35 40 45

Ser Cys He Ser Ser Ser Asp Gly He Thr Tyr Tyr Ala Asp Ser Val 50 55 60

Lys Gly Arg Phe Thr He Ser Arg Asp Asn Ser Lys Asn Thr Val Tyr 65 70 75 80

Leu Gin Met Asn Ser Leu Arg Pro Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Thr Asp Ser Gly Gly Tyr He Asp Tyr Asp Cys Met Gly Leu Gly 100 105 110

Tyr Asp Tyr Trp Gly Gin Gly Thr Leu Val Thr Val Ser Ser 115 120 125 <210> 133 <211> 126

<212> PRT <213> Artificial <220> <223> Mutated Lama sequence <400> 133

Glu val Gin Leu Val Glu Ser Gly Gly Gly Leu Val Gin Pro Gly Gly 15 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Ala Leu Asp Tyr Tyr 20 25 30

Ala He Gly Trp Phe Arg Gin Ala Pro Gly Lys Glu Arg Glu Gly Val 35 40 45

Ser Cys He Ser Ser Ser Gly Gly He Thr Tyr Tyr Val Asp Ser Val 50 55 60

Lys Gly Arg Phe Thr lie Ser Arg Asp Asn Ser Lys Asn Thr Val Tyr 65 70 75 80

Leu Gin Met Asn Ser Leu Arg Pro Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Thr Asp Ser Gly Gly Tyr lie Asp Tyr Asp Cys Met Gly Leu Gly 100 105 110

Tyr Asp Tyr Trp Gly Gin Gly Thr Leu Val Thr Val Ser Ser 115 120 125 <210> 134 <211> 126

<212> PRT <213> Artificial <220> <223> Mutated Lama sequence <400> 134

Glu Val Gin Leu Val Glu Ser Gly Gly Gly Leu Val Gin Pro Gly Gly 1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Ala Leu Asp Tyr Tyr 20 25 30

Ala He Gly Trp Phe Arg Gin Ala Pro Gly Lys Glu Arg Glu Gly Val 35 40 45

Ser Cys He Ser Ser Ser Gly Gly He Thr Tyr Tyr Ala Asp Ser Val 50 55 60

Lys Gly Arg Phe Thr He Ser Arg Asp Asn Ser Lys Asn Thr Val Tyr 65 70 75 80

Leu Gin Met Asn Ser Leu Arg Pro Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Thr Asp Ser Gly Gly Tyr lie Asp Tyr Asp Cys Met Gly Leu Gly 100 105 110

Tyr Asp Tyr Trp Gly Gin Gly Thr Leu Val Thr Val Ser Ser 115 120 125 <210> 135 <211> 126

<212> PRT <213> Artificial <220> <223> Mutated Lama sequence <400> 135

Asp Val Gin Leu Val Glu Ser Gly Gly Gly Leu Val Gin Pro Gly Gly 15 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Ala Leu Asp Tyr Tyr 20 25 30

Ala lie Gly Trp Phe Arg Gin Ala Pro Gly Lys Glu Arg Glu Gly Val 35 40 45

Ser Cys He Ser Ser Ser Gly Gly He Thr Tyr Tyr Ala Asp Ser Val 50 55 60

Lys Gly Arg Phe Thr He Ser Arg Asp Asn Ser Lys Asn Thr val Tyr 65 70 75 80

Leu Gin Met Asn Ser Leu Arg Pro Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Thr Asp Ser Gly Gly Tyr He Asp Tyr Asp Cys Met Gly Leu Gly 100 105 110

Tyr ASp Tyr Trp Gly Gin Gly Thr Leu Val thr Val Ser Ser 115 120 125 <210> 136 <211> 126

<212> PRT <213> Artificial <220> <223> Mutated Lama sequence <400> 136

Glu Val Gin Leu Val Glu Ser Gly Gly Gly Leu Val Gin Pro Gly Gly 15 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Ala Leu Asp Tyr Tyr 20 25 30

Ala lie Gly Trp Phe Arg Gin Ala Pro Gly Lys Glu Arg Glu Gly Val 35 40 45

Ser Cys He Ser Ser Ser Gly Gly He Thr Tyr Tyr Ala Asp Ser Val 50 55 60

Lys Gly Arg Phe Thr He Ser Arg Asp Asn Ser Lys Asn Thr Val Tyr 65 70 75 80

Leu Gin Met Asn Ser Leu Arg Pro Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Thr Asp Ser Gly Gly Tyr He Asp Tyr Asp Cys Gin Gly Leu Gly 100 105 110

Tyr Asp Tyr Trp Gly Gin Gly Thr Leu Val Thr Val Ser Ser 115 120 125 <210> 137 <211> 126

<212> PRT <213> Artificial <220> <223> Mutated Lama sequence <400> 137

Glu val Gin Leu Val Glu Ser Gly Gly Gly Leu Val Gin Pro Gly Gly 15 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Ala Leu Asp Tyr Tyr 20 25 30

Ala lie Gly Trp Phe Arg Gin Ala Pro Gly Lys Glu Arg Glu Gly Val 35 40 45

Ser Cys He Ser Ser Ser Gly Gly He Thr Tyr Tyr Ala Asp Ser Val 50 55 60

Lys Gly Arg Phe Thr He Ser Arg Asp Asn Ser Lys Asn Thr Val Tyr 65 70 75 80

Leu Gin Met Asn Ser Leu Arg Pro Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Thr Asp Ser Gly Gly Tyr He Asp Tyr Asp Cys Arg Gly Leu Gly 100 105 110

Tyr Asp Tyr Trp Gly Gin Gly Thr Leu Val Thr Val Ser Ser 115 120 125 <210> 138 <211> 126

<212> PRT <213> Artificial <220> <223> Mutated Lama sequence <400> 138

Glu Val Gin Leu Val Glu Ser Gly Gly Gly Leu Val Gin Pro Gly Gly 1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Ala Leu Asp Tyr Tyr 20 25 30

Ala He Gly Trp Phe Arg Gin Ala Pro Gly Lys Glu Arg Glu Gly Val 3r /in /iK

Ser Cys lie Ser Ser Ser Gly Gly lie Thr Tyr Tyr Ala Asp Ser Val 50 55 60

Lys Gly Arg Phe Thr lie Ser Arg Asp Asn Ser Lys Asn Thr Val Tyr 65 70 75 80

Leu Gin Met Asn Ser Leu Arg Pro Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Thr Asp Ser Gly Gly Tyr He Asp Tyr Asp Cys Ser Gly Leu Gly 100 105 HO

Tyr Asp Tyr Trp Gly Gin Gly Thr Leu Val Thr Val Ser Ser 115 120 125 <210> 139 <211> 126

<212> PRT <213> Artificial <220> <223> Mutated Lama sequence <400> 139

Glu Val Gin Leu Val Glu Ser Gly Gly Gly Leu Val Gin Pro Gly Gly 15 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Ala Leu Asp Tyr Tyr 20 25 30

Ala lie Gly Trp Phe Arg Gin Ala Pro Gly Lys Glu Arg Glu Gly Val 35 40 45

Ser Cys He Ser Ser Ser Gly Gly He Thr Tyr Tyr Ala Asp Ser Val 50 55 60

Lys Gly Arg Phe Thr He Ser Arg Asp Asn Ser Lys Asn Thr Val Tyr 65 70 75 80

Leu Gin Met Asn Ser Leu Arg Pro Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Thr Glu Ser Gly Gly Tyr He Asp Tyr Asp Cys Gin Gly Leu Gly 100 105 110

Tyr Asp Tyr Trp Gly Gin Gly Thr Leu Val Thr Val Ser Ser 115 120 125 <210> 140 <211> 126

<212> PRT <213> Artificial <220> <223> Mutated Lama sequence <400> 140

Glu Val Gin Leu Val Glu Ser Gly Gly Gly Leu Val Gin Pro Gly Gly 1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Ala Leu Asp Tyr Tyr 20 25 30

Ala lie Gly Trp Phe Arg Gin Ala Pro Gly Lys Glu Arg Glu Gly Val 35 40 45

Ser Cys He Ser Ser Ser Gly Gly He Thr Tyr Tyr Ala Asp Ser Val 50 55 60

Lys Gly Arg Phe Thr He Ser Arg Asp Asn Ser Lys Asn Thr Val Tyr 65 70 75 80

Leu Gin Met ASrt Ser Leu Arg Pro Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Thr Glu Ser Gly Gly Tyr lie Asp Tyr Asp Cys Arg Gly Leu Gly 100 105 110

Tyr Asp Tyr Trp Gly Gin Gly Thr Leu Val Thr Val Ser Ser 115 120 125 <210> 141 <211> 126

<212> PRT <213> Artificial <220> <223> Mutated Lama sequence <400> 141

Glu Val Gin Leu Val Glu Ser Gly Gly Gly Leu Val Gin Pro Gly Gly 15 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Ala Leu Asp Tyr Tyr 20 25 30

Ala He Gly Trp Phe Arg Gin Ala Pro Gly Lys Glu Arg Glu Gly Val 35 40 45

Ser Cys lie Ser Ser Ser Gly Gly He Thr Tyr Tyr Ala Asp Ser Val 50 55 60

Lys Gly Arg Phe Thr He Ser Arg Asp Asn Ser Lys Asn Thr Val Tyr 65 70 75 80

Leu Gin Met Asn Ser Leu Arg Pro Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Thr Glu Ser Gly Gly Tyr lie Asp Tyr Asp Cys Ser Gly Leu Gly 100 105 110

Tyr Asp Tyr Trp Gly Gin Gly Thr Leu Val Thr Val Ser Ser 115 120 125 <210> 142 <211> 129

<212> PRT <213> Artificial <220> <223> Mutated Lama sequence <400> 142

Glu Val Gin Leu Val Glu Ser Gly Gly Gly Leu Val Gin Ala Gly Gly 15 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Leu Asp Asp Tyr 20 25 30

Ala He Gly Trp Phe Arg Gin Ala Pro Gly Lys Glu Arg Glu Gly Val 35 40 45

Ser Thr He Arg Asp Ser Asp Gly Ser Thr Tyr Tyr Ala Asp Ser Val 50 55 60

Lys Gly Arg Phe Thr He Ser Ser Asp Asn Asp Lys Asn Thr Val Tyr 65 70 75 80

Leu Gin Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Ala Val Pro Ala Gly Arg Leu Arg Phe Gly Glu Gin Trp Tyr Pro 100 105 110

Leu Tyr Glu Tyr Asp Ala Trp Gly Gin Gly Thr Leu Val Thr Val Ser 115 120 125

Ser <210> 143 <211> 129

<212> PRT <213> Artificial <220> <223> Mutated Lama sequence <400> 143

Glu Val Gin Leu Val Glu Ser Gly Gly Gly Leu Val Gin Ala Gly Gly 15 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Leu Asp Asp Tyr 20 25 30

Ala lie Gly Trp Phe Arg Gin Ala Pro Gly Lys Glu Arg Glu Gly Val 35 40 45

Ser Ala lie Arg Asp Ser Asp Gly Ser Thr Tyr Tyr Ala Asp Ser Val 50 55 60

Lys Gly Arg Phe Thr He Ser Ser Asp Asn Asp Lys Asn Thr Val Tyr 65 70 75 80

Leu Gin Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Ala Val Pro Ala Gly Arg Leu Arg Phe Gly Glu Gin Trp Tyr Pro 100 105 110

Leu Tyr Glu Tyr Asp Ala Trp Gly Gin Qly Thr Leu Val Thr Val Ser 115 120 125

Ser <210> 144 <211> 129

<212> PRT <213> Artificial <220> <223> Mutated Lama sequence <400> 144

Glu Val Gin Leu Val Glu Ser Gly Gly Gly Leu Val Gin Ala Gly Gly 1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr LeU Asp Asp Tyr 20 25 30

Ala He Gly Trp Phe Arg Gin Ala Pro Gly Lys Glu Arg Glu Gly Val 35 40 45

Ser Ser He Arg Asp Asn Asp Gly Ser Thr Tyr Tyr Ala Asp Ser Val 50 55 60

Lys Gly Arg Phe Thr He Ser Ser Asp Asn Asp Lys Asn Thr Val Tyr 65 70 75 80

Leu Gin Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Ala Val Pro Ala Gly Arg Leu Arg Phe Gly Glu Gin Trp Tyr Pro 100 105 110

Leu Tyr Glu Tyr Asp Ala Trp Gly Gin Gly Thr Leu Val Thr Val Ser 115 120 125

Ser <210> 145 <211> 129

<212> PRT <213> Artificial <220> <223> Mutated Lama sequence <400> 145

Glu Val Gin Leu val Glu Ser Gly Gly Gly Leu Val Gin Pro Gly Gly 1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Leu Asp Asp Tyr 20 25 30

Ala He Gly Trp Phe Arg Gin Ala Pro Gly Lys Glu Arg Glu Gly Val 35 40 45

Ser Ser He Arg Asp Asn Asp Gly Ser Thr Tyr Tyr Ala Asp Ser Val 50 55 60

Lys Gly Arg Phe Thr He Ser Ser Asp Asn Ala Lys Asn Thr val Tyr 65 70 75 80

Leu Gin Met Asn Ser Leu Arg Pro Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Ala Val Pro Ala Gly Arg Leu Arg Phe Gly Glu Gin Trp Tyr Pro 100 105 110

Leu Tyr Glu Tyr Asp Ala Trp Gly Gin Gly Thr Leu Val Thr Val Ser 115 120 125

Ser <210> 146 <211> 129

<212> PRT <213> Artificial <220> <223> Mutated Lama sequence <400> 146

Glu Val Gin Leu Val Glu Ser Gly Gly Gly Leu Val Gin Pro Gly Gly 15 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Leu Asp Asp Tyr zu zt> ju

Ala He Gly Trp Phe Arg Gin Ala Pro Gly Lys Glu Arg Glu Gly Val 35 40 45

Ser Ser He Arg Asp Asn Asp Gly Ser Thr Tyr Tyr Ala Asp Ser Val 50 55 60

Lys Gly Arg Phe Thr He Ser Ser Asp Asn Ser Lys Asn Thr Val Tyr 65 70 75 80

Leu Gin Met Asn Ser Leu Arg Pro Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Ala Val Pro Ala Gly Arg Leu Arg Phe Gly Glu Gin Trp Tyr Pro 100 105 110

Leu Tyr Glu Tyr Asp Ala Trp Gly Gin Gly Thr Leu Val Thr Val Ser 115 120 125

Ser <210> 147 <211> 129

<212> PRT <213> Artificial <220> <223> Mutated Lama sequence <400> 147

Glu Val Gin Leu Val Glu Ser Gly Gly Gly Leu Val Gin Pro Gly Gly 15 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Leu Asp Asp Tyr 20 25 30

Ala He Gly Trp Phe Arg Gin Ala Pro Gly Lys Glu Arg Glu Gly Val 35 40 45

Ser Ser He Arg Asp Asn Asp Gly Ser Thr Tyr Tyr Ala Asp Ser Val 50 55 60

Lys Gly Arg Phe Thr He Ser Arg Asp Asn Ala Lys Asn Thr Val Tyr 65 70 75 80

Leu Gin Met Asn Ser Leu Arg Pro Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Ala Val Pro Ala Gly Arg Leu Arg Phe Gly Glu Gin Trp Tyr Pro 100 105 110

Leu Tyr Glu Tyr Asp Ala Trp Gly Gin Gly Thr Leu Val Thr Val Ser 115 120 125

Ser <210> 148 <211> 129

<212> PRT <213> Artificial <220> <223> Mutated Lama sequence <400> 148

Glu Val Gin Leu Val Glu Ser Gly Gly Gly Leu Val Gin Ala Gly Gly 15 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Leu Asp Asp Tyr 20 25 30

Ala lie Gly Trp Phe Arg Gin Ala Pro Gly Lys Glu Arg Glu Gly Val 35 40 45

Ser Ser He Arg Asp Ser Asp Gly Ser Thr Tyr Tyr Ala Asp Ser Val 50 55 60

Lys Gly Arg Phe Thr He Ser Ser Asp Asn Asp Lys Asn Thr val Tyr 65 70 75 80

Leu Gin Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Ala Val Pro Ala Gly Arg Leu Arg Phe Gly Glu Gin Trp Tyr Pro 100 105 110

Leu Tyr Glu Tyr Asp Ala Trp Gly Gin Gly Thr Leu val Thr Val Ser 115 120 125

Ser <210> 149 <211> 129

<212> PRT <213> Artificial <220> <223> Mutated Lama sequence <400> 149

Glu Val Gin LeU Val Glu Ser Gly Gly Gly Leu Val Gin Pro Gly Gly 15 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Leu Asp Asp Tyr 20 25 30 &amp;la Ti e /21 *r Tw* Dbe JLrvr /21 τί 2k 1 a Drc /21 ir T.we /21 ii 2kr»rr /21 π /21 vr Val cxx-a. -i- x.^ wx._y xj_£> x ns nxa x j-t_s wx.^ xj^ <9 w-i-<x x-xj-k^ vxu wx-jr vex 35 40 45

Ser Ser lie Arg Asp Asn Asp Gly Ser Thr Tyr Tyr Ala Asp Ser val 50 55 60

Lys Gly Arg Phe Thr He Ser Ser Asp Asn Asp Lys Asn Thr Val Tyr 65 70 75 80

Leu Gin Met Asn Ser Leu Arg Pro Glu Asp Thr Ala Val Tyr Tyr Gys 85 90 95

Ala Ala Val Pro Ala Gly Arg Leu Arg Phe Gly Glu Gin Trp Tyr Pro 100 105 110

Leu Tyr Glu Tyr Asp Ala Trp Gly Gin Gly Thr Leu Val Thr Val Ser 115 120 125

Ser <210> 150 <211> 129

<212> PRT <213> Artificial <220> <223> Mutated Lama sequence <400> 150

Glu Val Gin Leu Val Glu Ser Gly Gly Gly Leu Val Gin Pro Gly Gly 1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Leu Asp Asp Tyr 20 25 30

Ala He Gly Trp Phe Arg Gin Ala Pro Gly Lys Glu Arg Glu Gly Val 35 40 45

Ser Ser He Arg Asp Asn Asp Gly Ser Thr Tyr Tyr Ala Asp Ser Val 50 55 60

Lys Gly Arg Phe Thr He Ser Arg Asp Asn Asp Lys Asn Thr Val Tyr 65 70 75 80

Leu Gin Met Asn Ser Leu Arg Pro Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Ala Val Pro Ala Gly Arg Leu Arg Phe Gly Glu Gin Trp Tyr Pro 100 105 110

Leu Tyr Glu Tyr Asp Ala Trp Gly Gin Gly Thr Leu Val Thr Val Ser 115 120 125

Ser <210> 151 <211> 129

<212> PRT <213> Artificial <220> <223> Mutated Lama sequence <400> 151

Glu Val Gin Leu Val Glu Ser Gly Gly Gly Leu Val Gin Pro Gly Gly 15 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Leu Asp Asp Tyr 20 25 30

Ala lie Gly Trp Phe Arg Gin Ala Pro Gly Lys Glu Arg Glu Gly Val 35 40 45

Ser Ser He Arg Asp Asn Asp Gly Ser Thr Tyr Tyr Ala Asp Ser Val 50 55 60

Lys Gly Arg Phe Thr He Ser Arg Asp Asn Ser Lys Asn Thr Val Tyr 65 70 75 80

Leu Gin Met Asn Ser Leu Arg Pro Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Ala Val Pro Ala Gly Arg Leu Arg Phe Gly Glu Gin Trp Tyr Pro 100 105 110

Leu Tyr Glu Tyr Asp Ala Trp Gly Gin Gly Thr Leu Val Thr Val Ser 115 120 125

Ser <210> 152 <211> 129

<212> PRT <213> Artificial <220> <223> Mutated Lama sequence <400> 152

Glu val Gin Leu Val Glu Ser Gly Gly Gly Leu Val Gin Ala Gly Gly 15 10 15

Ser Leu Arg LeU Ser Cys Ala Ala Ser Gly Phe Thr Leu Asp Asp Tyr 20 25 30

Ala He Gly Trp Phe Arg Gin Ala Pro Gly Lys Glu Arg Glu Gly Val 35 40 45

Sei Spr He Aro Asn Asn Rlv Glv Ser Thr Tv' Tvr Ala Asn Ser Val — —— — — — — — — — J ---J — - — — -J— -J— £T — —' — · — — 50 55 60

Lys Gly Arg Phe Thr He Ser Ser Asp Asn Asp Lys Asn Thr Val Tyr 65 70 75 80

Leu Gin Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Ala Val Pro Ala Gly Arg Leu Arg Phe Gly Glu Gin Trp Tyr Pro 100 105 110

Leu Tyr Glu Tyr Asp Ala Trp Gly Gin Gly Thr Leu Val Thr Val Ser 115 120 125

Ser <210> 153 <211> 129

<212> PRT <213> Artificial <220> <223> Mutated Lama sequence <400> 153

Glu Val Gin Leu Val Glu Ser Gly Gly Gly Leu Val Gin Ala Gly Gly 1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Leu Asp Asp Tyr 20 25 30

Ala He Gly Trp Phe Arg Gin Ala Pro Gly Lys Glu Arg Glu Gly Val 35 40 45

Ser Ala He Arg Asp Asn Asp Gly Ser Thr Tyr Tyr Ala Asp Ser Val 50 55 60

Lys Gly Arg Phe thr He Ser Ser Asp Asn Asp Lys Asn Thr Val Tyr 65 70 75 80

Leu Gin Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Ala Val Pro Ala Gly Arg Leu Arg Phe Gly Glu Gin Trp Tyr Pro 100 105 110

Leu Tyr Glu Tyr Asp Ala Trp Gly Gin Gly Thr Leu Val Thr Val Ser 115 120 125

Ser <210> 154 <211> 129

<212> PRT <213> Artificial <220> <223> Mutated Lama sequence <400> 154

Glu Val Gin Leu Val Glu Ser Gly Gly Gly Leu Val Gin Ala Gly Gly 15 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Leu Asp Asp Tyr 20 25 30

Ala lie Gly Trp Phe Arg Gin Ala Pro Gly Lys Glu Arg Glu Gly Val 35 40 45

Ser Ser lie Arg Asp He Asp Gly Ser Thr Tyr Tyr Ala Asp Ser Val 50 55 60

Lys Gly Arg Phe Thr He Ser Ser Asp Asn Asp Lys Asn Thr Val Tyr 65 70 75 80

Leu Gin Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Ala Val Pro Ala Gly Arg Leu Arg Phe Gly Glu Gin Trp Tyr Pro 100 105 110

Leu Tyr Glu Tyr Asp Ala Trp Gly Gin Gly Thr Leu Val Thr Val Ser 115 120 125

Ser <210> 155 <211> 129

<212> PRT <213> Artificial <220> <223> Mutated Lama sequence <400> 155

Glu Val Gin Leu Val Glu Ser Gly Gly Gly Leu Val Gin Ala Gly Gly 1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Val Ser Gly Phe Thr Leu Asp Asp Tyr 20 25 30

Ala He Gly Trp Phe Arg Gin Ala Pro Gly Lys Glu Arg Glu Gly Val 35 40 45

Ser Ser He Arg Asp Asn Asp Gly Ser Thr Tyr Tyr Ala Asp Ser Val 50 55 60

Lys Gly Arg Phe Thr lie Ser Ser Asp Asn Asp Lys Asn Thr Val Tyr 65 70 75 80

Leu Gin Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Ala Val Pro Ala Gly Arg Leu Arg Phe Gly Glu Gin Trp Tyr Pro 100 105 110

Leu Tyr Glu Tyr Asp Ala Trp Gly Gin Gly Thr Leu Val Thr Val Ser 115 120 125

Ser <210> 156 <211> 129

<212> PRT <213> Artificial <220> <223> Mutated Lama sequence <400> 156

Glu Val Gin Leu Val Glu Ser Gly Gly Gly Leu Val Gin Ala Gly Gly 1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Leu Asp Asp Tyr 20 25 30

Ala lie Gly Trp Phe Arg Gin Ala Pro Gly Lys Glu Arg Glu Gly Val 35 40 45

Ser Ala lie Arg Asp Ser Asp Gly Ser Thr Tyr Tyr Ala Asp Ser Val 50 55 60

Lys Gly Arg Phe Thr He Ser Ser Asp Asn Asp Lys Asn Thr Val Tyr 65 70 75 80

Leu Gin Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Ala Val Pro Ala Gly Arg Leu Arg Phe Gly Glu Gin Trp Tyr Pro 100 105 110

Leu Tyr Glu Tyr Asp Ala Trp Gly Gin Gly Thr Leu Val Thr Val Ser 115 120 125

Ser <210> 157 <211> 129

<212> PRT <213> Artificial <220> <223> Mutated Lama sequence <400> 157

Glu Val Gin Leu Val Glu Ser Gly Gly Gly Leu Val Gin Ala Gly Gly 1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Leu Asp Asp Tyr 20 25 30

Ala lie Gly Trp Phe Arg Gin Ala Pro Gly Lys Glu Arg Glu Gly Val 35 40 45

Ser Ala He Arg Asp Ser Gly Gly Ser Thr Tyr Tyr Ala Asp Ser Val 50 55 60

Lys Gly Arg Phe Thr He Ser Ser Asp Asn Asp Lys Asn Thr Val Tyr 65 70 75 80

Leu Gin Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Ala Val Pro Ala Gly Arg Leu Arg Phe Gly Glu Gin Trp Tyr Pro 100 105 110

Leu Tyr Glu Tyr Asp Ala Trp Gly Gin Gly Thr Leu Val Thr Val Ser 115 120 125

Ser <210> 158 <211> 129

<212> PRT <213> Artificial <220> <223> Mutated Lama sequence <400> 158

Glu Val Gin Leu Val Glu Ser Gly Gly Gly Leu Val Gin Pro Gly Gly 15 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Leu Asp Asp Tyr 20 25 30

Ala He Gly Trp Phe Arg Gin Ala Pro Gly Lys Glu Arg Glu Gly Val 35 40 45

Ser Ala He Arg Asp Asn Gly Gly Ser Thr Tyr Tyr Ala Asp Ser Val 50 55 60

Lys Gly Arg Phe Thr He Ser Ser Asp Asn Ser Lys Asn Thr Val Tyr 65 70 75 80

Leu Gin Met Asn Ser Leu Arg Pro Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Ala Val Pro Ala Gly Arg Leu Arg Phe Gly Glu Gin Trp Tyr Pro 100 105 110

Leu Tyr Glu Tyr Asp Ala Trp Gly Gin Gly Thr Leu Val Thr Val Ser 115 120 125

Ser <210> 159 <211> 129

<212> PRT <213> Artificial <220> <223> Mutated Lama sequence <400> 159

Asp Val Gin Leu Val Glu Ser Gly Gly Gly Leu Val Gin Pro Gly Gly 1 5 10 15

Ser Leu Arg Leu Ser CyS Ala Ala Ser Gly Phe Thr Leu Asp Asp Tyr 20 25 30

Ala lie Gly Trp Phe Arg Gin Ala Pro Gly Lys Glu Arg Glu Gly Val 35 40 45

Ser Ala He Arg Asp Asn Gly Gly Ser Thr Tyr Tyr Ala Asp Ser Val 50 55 60

Lys Gly Arg Phe Thr He Ser Ser Asp Asn Ser Lys Asn Thr Val Tyr 65 70 75 80

Leu Gin Met Asn Ser Leu Arg Pro Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Ala Val Pro Ala Gly Arg Leu Arg Phe Gly Glu Gin Trp Tyr Pro 100 105 110

Leu Tyr Glu Tyr Asp Ala Trp Gly Gin Gly Thr Leu Val Thr Val Ser 115 120 125

Ser <210> 160 <211> 129

<212> PRT <213> Artificial <220> <223> Mutated Lama sequence <400> 160

Glu Val Gin Leu Val Glu Ser Gly Gly Gly Leu Val Gin Pro Gly Gly 15 10 15

Ser Leu Arg Leu Ser Cys Ala Val Ser Gly lie Thr Leu Asp Asp Tyr 20 25 30

Ala He Gly Trp Phe Arg Gin Ala Pro Gly Lys Glu Arg Glu Gly Val 35 40 45

Ser Ser He Arg Asp Asn Gly Gly Ser Thr Tyr Tyr Ala Asp Ser Val 50 55 60

Lys Gly Arg Phe Thr He Ser Ser Asp Asn Ser Lys Asn Thr Val Tyr 65 70 75 80

Leu Gin Met Asn Ser Leu Arg Pro Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Ala Val Pro Ala Gly Arg Leu Arg Tyr Gly Glu Gin Trp Tyr Pro 100 105 110

He Tyr Glu Tyr Asp Ala Trp Gly Gin Gly Thr Leu Val Thr Val Ser 115 120 125

Ser <210> 161 <211> 129

<212> PRT <213> Artificial <220> <223> Mutated Lama sequence <400> 161

Glu Val Gin Leu Val Glu Ser Gly Gly Gly Leu Val Gin Pro Gly Gly 15 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly He Thr Leu Asp Asp Tyr 20 25 30

Ala He Gly Trp Phe Arg Gin Ala Pro Gly Lys Glu Arg Glu Gly Val 35 40 45

Ser Ala He Arg Asp Asn Gly Gly Ser Thr Tyr Tyr Ala Asp Ser Val 50 55 60

Lys Gly Arg Phe Thr He Ser Ser Asp Asn Ser Lys Asn Thr Val Tyr 65 70 75 80

Leu Gin Met Asn Ser Leu Arg Pro Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Ala Val Pro Ala Gly Arg Leu Arg Tyr Gly Glu Gin Trp Tyr Pro 100 105 110 lie Tyr Glu Tyr Asp Ala Trp Gly Gin Gly Thr Leu Val Thr Val Ser 115 120 125

Ser <210> 162 <211> 129

<212> PRT <213> Artificial <220> <223> Mutated Lama sequence <400> 162

Glu Val Gin Leu Val Glu Ser Gly Gly Gly Leu Val Gin Pro Gly Gly 15 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly lie Thr Leu Asp Asp Tyr 20 25 30

Ala He Gly Trp Phe Arg Gin Ala Pro Gly Lys Glu Arg Glu Gly Val 35 40 45

Ser Ala He Arg Glu Ser Gly Gly Ser Thr Tyr Tyr Ala Asp Ser Val 50 55 60

Lys Gly Arg Phe Thr lie Ser Ser Asp Asn Ser Lys Asn Thr Val Tyr 65 70 75 80

Leu Gin Met Asn Ser Leu Arg Pro Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Ala Val Pro Ala Gly Arg Leu Arg Tyr Gly Glu Gin Trp Tyr Pro 100 105 110 lie Tyr Glu Tyr Asp Ala Trp Gly Gin Gly Thr Leu Val Thr Val Ser 115 120 125

Ser <210> 163 <211> 129

<212> PRT <213> Artificial <220> <223> Mutated Lama sequence <400> 163

Glu Val Gin Leu Val Glu Ser Gly Gly Gly Leu Val Gin Pro Gly Gly 15 10 15 ser Leu Arg Leu ser cys Ala A±a ser my lie inr Leu Asp Asp Tyr 20 25 30

Ala lie Gly Trp Phe Arg Gin Ala Pro Gly Lys Glu Arg Glu Gly Val 35 40 45

Ser Ala He Arg Ser Ser Gly Gly Ser Thr Tyr Tyr Ala Asp Ser Val 50 55 60

Lys Gly Arg Phe Thr He Ser Ser Asp Asn Ser Lys Asn Thr Val Tyr 65 70 75 80

Leu Gin Met Asn Ser Leu Arg Pro Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Ala Val Pro Ala Gly Arg Leu Arg Tyr Gly Glu Gin Trp Tyr Pro 100 105 110

He Tyr Glu Tyr Asp Ala Trp Gly Gin Gly Thr Leu Val Thr Val Ser 115 120 125

Ser <210> 164 <211> 129

<212> PRT <213> Artificial <220> <223> Mutated Lama sequence <400> 164

Glu Val Gin Leu Val Glu Ser Gly Gly Gly Leu Val Gin Pro Gly Gly 15 10 15

Ser Leu Arg Leu Ser Cys Ala Val Ser Gly He Thr Leu Asp Asp Tyr 20 25 30

Ala He Gly Trp Phe Arg Gin Ala Pro Gly Lys Glu Arg Glu Gly Val 35 40 45

Ser Ala He Arg Asp Asn Gly Gly Ser Thr Tyr Tyr Ala Asp Ser Val 50 55 60

Lys Gly Arg Phe Thr He Ser Ser Asp Asn Ser Lys ASn Thr Val Tyr 65 70 75 80

Leu Gin Met Asn Ser Leu Arg Pro Glu Asp Thr Ala val Tyr Tyr Cys 85 90 95

Ala Ala Val Pro Ala Gly Arg Leu Arg Tyr Gly Glu Gin Trp Tyr Pro 100 105 110

He Tyr Glu Tyr Asp Ala Trp Gly Gin Gly Thr Leu Val Thr Val Ser 115 120 125

Ser <210> 165 <211> 129

<212> PRT <213> Artificial <220> <223> Mutated Lama sequence <400> 165

Glu Val Gin Leu Val Glu Ser Gly Gly Gly Leu Val Gin Pro Gly Gly 1 5 10 15

Ser Leu Arg Leu Ser Cys Ala Val Ser Gly lie Thr Leu Asp Asp Tyr 20 25 30

Ala He Gly Trp Phe Arg Gin Ala Pro Gly Lys Glu Arg Glu Gly Val 35 40 45

Ser Ala He Arg Glu Ser Gly Gly Ser Thr Tyr Tyr Ala Asp Ser Val 50 55 60

Lys Gly Arg Phe Thr He Ser Ser Asp Asn Ser Lys Asn Thr Val Tyr 65 70 75 80

Leu Gin Met Asn Ser Leu Arg Pro Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Ala Val Pro Ala Gly Arg Leu Arg Tyr Gly Glu Gin Trp Tyr Pro 100 105 110

He Tyr Glu Tyr Asp Ala Trp Gly Gin Gly Thr Leu Val Thr Val Ser 115 120 125

Ser <210> 166 <211> 129

<212> PRT <213> Artificial <220> <223> Mutated Lama sequence <400> 166

Glu Val Gin Leu Val Glu Ser Gly Gly Gly Leu Val Gin Pro Gly Gly 15 10 15

Ser Leu Arg LeU Ser Cys Ala Val Ser Gly He Thr Leu Asp Asp Tyr 20 25 30

Ala lie Gly Trp Phe Arg Gin Ala Pro Gly Lys Glu Arg Glu Gly Val 35 40 45

Ser Ala lie Arg Ser Ser Gly Gly Ser Thr Tyr Tyr Ala Asp Ser Val 50 55 60

Lys Gly Arg Phe Thr He Ser Ser Asp Asn Ser Lys Asn Thr Val Tyr 65 70 75 80

Leu Gin Met Asn Ser Leu Arg Pro Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Ala Val Pro Ala Gly Arg Leu Arg Tyr Gly Glu Gin Trp Tyr Pro 100 105 110

He Tyr Glu Tyr Asp Ala Trp Gly Gin Gly Thr Leu Val Thr Val Ser 115 120 125

Ser <210> 167 <211> 129

<212> PRT <213> Artificial <220> <223> Mutated Lama sequence <400> 167

Glu Val Gin Leu Val Glu Ser Gly Gly Gly Leu Val Gin Pro Gly Gly 15 10 15

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Leu Asp Asp Tyr 20 25 30

Ala He Gly Trp Phe Arg Gin Ala Pro Gly Lys Glu Arg Glu Gly Val 35 40 45

Ser Ala He Arg Ser Ser Gly Gly Ser Thr Tyr Tyr Ala Asp Ser Val 50 55 60

Lys Gly Arg Phe Thr He Ser Ser Asp Asn Ser Lys Asn Thr Val Tyr 65 70 75 80

Leu Gin Met Asn Ser Leu Arg Pro Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95

Ala Ala Val Pro Ala Gly Arg Leu Arg Phe Gly Glu Gin Trp Tyr Pro 100 105 110

Leu Tyr Glu Tyr Asp Ala Trp Gly Gin Gly Thr Leu Val Thr Val Ser 115 120 125

Ser <210> 168 <211> 5

<212> PRT <213> Artificial <220> <223> Mutated Lama sequence <400> 168

Asp Tyr Ala lie Gly 1 5 <210> 169 <211> 5

<212> PRT <213> Artificial <220> <223> Mutated Lama sequence <400> 169

Asp Tyr Ala Leu Gly 1 5 <210> 170 <211> 5

<212> PRT <213> Artificial <220> <223> Mutated Lama sequence <400> 170

Tyr Tyr Ala He Gly 1 5 <210> 171 <211> 17

<212> PRT <213> Artificial <220> <223> Mutated Lama sequence <400> 171

Ala lie Arg Ser Ser Gly Gly Ser Thr Tyr Tyr Ala Asp Ser Val Lys 15 10 15

Gly <210> 172 <211> 17

<212> PRT <213> Artificial <220> <223> Mutated Lama sequence <400> 172

Cys He Arg Cys Ser Gly Gly Ser Thr Tyr Tyr Ala Asp Ser Val Lys 1 5 10 15

Gly <21O> 173 <211> 17

<212> PRT <213> Artificial <220> <223> Mutated Lama sequence <400> 173

Cys He Ser Ser Ser Gly Gly He Thr Tyr Tyr Ala Asp Ser Val Lys 15 10 15

Gly <21 O> 174 <211> 20

<212> PRT <213> Artificial <220> <223> Mutated Lama sequence <400> 174

Val Pro Ala Gly Arg Leu Arg Phe Gly Glu Gin Trp Tyr Pro Leu Tyr 15 10 15

Glu Tyr Asp Ala 20 <210> 175 <211> 20

<212> PRT <213> Artificial <220> <223> Mutated Lama sequence <400> 175

Val Pro Ala Gly Arg Leu Arg Tyr Gly Glu Gin Trp Tyr Pro He Tyr 15 10 15

Glu Tyr Asp Ala 20 <210> 176 <211> 15

<212> PRT <213> Artificial <220> <223> Mutated Lama sequence <400> 176

Ser lie Val Pro Arg Ser Lys Leu Glu Pro Tyr Glu Tyr Asp Ala 15 10 15 <210> 177 <211> 17

<212> PRT <213> Artificial <220> <223> Mutated Lama sequence <400> 177

Asp Ser Gly Gly Tyr lie Asp Tyr Asp Cys Ser Gly Leu Gly Tyr Asp 15 10 15

Tyr <210> 178 <211> 387

<212> DNA <213> Artificial <220> <223> Mutated Lama sequence <400> 178 gaggtacagc tggtcgagtc aggtggcgga ttagtgcagc ctgggggttc tctgcgctta 60 tcttgtgceg catcaggott cacactggat gactacgcca tcggctggtt ccggcaagcg 120 cctggaaaag aacgcgaagg tgtttcagca atccgttcaa gcggtggttc aacatattac 180 gccgactetg ttaaaggacg cttcaccatt agctccgaca atagtaaaaa tacagtctac 240 ttacaaatga acagtttacg cccagaagat actgcggtat actattgcgc tgccgtgoct 300 gctggtcgct tacgctttgg cgagcaatgg tatcctctgt acgagtacga cgcctgggga 360 cagggtacgc tggtaacggt ttcaagc 387 <210> 179 <211> 387

<212> DNA <213> Artificial <220> <223> Mutated Lama sequence <400> 179 gaagtgcaac tggttgagtc aggtggcgga ctggtgcaac cgggtggttc actgcgcctg 60 agttgcgcag ttagcggtat taccctggat gattatgcaa ttggttggtt tcgqcaagcc 120 ccaggcaaag agcgtgaagg cgttagcgca attcgtagca gcggtggtag cacctattat 180 gccgattoag ttaaaggccg ttttacgatc agcagcgata acagtaaaaa cacggtttat 240 ctgcaaatga actcattacg tccagaggac actgcagttt actattgcgc agcagttccg 300 gcaggtcgtc tgcgttatgg tgaacagtgg tatccgattt atgaatatga tgcatggggt 360 caaggtacac tggttacagt gagtagc 387 <210> 180 <211> 372

<212> DNA <213> Artificial <220> <223> Mutated Lama sequence <400> 180 gaagtgcagt tagtcgaaag tggcggaggc ctggtacaac ctggtggeag tctgcgctta 60 tcttgtgccg cttcaggttt tacattcgac gactacgccc tggggtggtt ccggcaagcg 120 cctggaaaag aacgtgaggg cgtttcatgc attcgttgtt caggtggttc aacctattat 180 gccgatagtg taaaaggtcg gttcaccatt agtagcgaca atagcaagaa tacagtctat 240 ctgcaaatga actctttacg tcctgaagat actgcggtgt actactgcgc tgcatcaatc 300 gttcctcgtt Caaaacttga accttacgag tacgacgcct ggggtcaggg tacgttagta 360 acggtgtcaa gc 372 <210> 181 <211> 378

<212> DNA <213> Artificial <220> <223> Mutated Lama sequence <400> 181 gaagtgcaac tggttgagtc aggtggcggt ttagtgCaaC cgggtggttc gctgCgCctg 60 agttgcgcag ccagcggttt tgcactggat tattatgcaa ttggttggtt tcgccaagcc 120 ccaggcaaag agcgtgaagg cgttagctgt attagcagca gcggtggtat tacctattat 180 gccgattcag ttaaaggccg ttttacgatc tctcgtgata atagtaaaaa cacggtttac 240 ctgcagatga actcattaag accagaggac actgcagttt aatattgtga aaccgatagc 300 ggtggctata ttgattatga ttgtagcggt ctgggctacg attattgggg acaaggtacg 360 ctggtgacag ttagcagc 378 <210> 182 <211> 3375

<212> DNA <213> Homo sapiens <400> 182 atggactctt tagccagctt agttctctgt ggagtaagat tgctcctttc tggaactgtg 60 gaaggtgcca tggacttgat cttgatcaat tccctacctc ttgtatntga tgctgaaaca 120 tatataaaat gcattgcctc tgggtggcgc ccccatgagc ggataaaaat aggaagggac 180 tttgaagqet taatgaaeaa gcaccaggat ccgctggaag ttaqtqaaga tgtgaqaaga 240 gaatgggcta aaaaagttgt ttggaagaga gaaaaggcta gtaagatcaa tggtgcttat 300 ttctgtgaag ggqgagttag aggagaggca ataaggataa gaaccatgaa gatgcgtcaa 360 caagcttcct tcctaccagc taatttaaet atgactgtgg acaagggaga taaagtgaae 420 atatctttca aaaaggtatt gattaaagaa gaagatgcag tgatttacaa aaatggttaa 480 ttaatggatt cagtgccccg gcatgaagta cctgatattc tagaagtaca cctgcctcat 540 gctcagcccc aggatgctgg agtgtactcg gccaggtata taggaggaaa cctcttcacc 600 tcggccttca ccaggctgat agtccggaga tgtgaagccc agaagtgggg acctgaatgc 660 aaccatctct gtactgcttg tatgaacaat ggtgtctgcc atgaagatac tggagaatgc 720 atttgccctc ctgggtttat gggaaggacg tgtgagaagg cttgtgaact gcacacgttt 780 ggcagaactt gtaaagaaag gtgcagtgga caagagggat gcaagtctta tgtgttctgt 840 ctccctgacc cctatgggtg ttcctgtgcc acaggotgga agggtctgca gtgcaatgaa 900 gcatgccacc ctggttttta cgggccagat tgtaagctta ggtgoagctg caaoaatggg 960 gagatgtgtg atcgcttcca aggatgtctc tgctctccag gatggcaggg gctccagtgt 1020 gagagagaag goataccgag gatgacccca aagatagtgg atttgccaga tcatatagaa 1080 gtaaacagtg gtaaatttaa tcccatttgc aaagcttctg gctggccgct acctactaat 1140 gaagaaatga ccetggtgaa gccggatggg acagtgctcc atcqaaaaga ctttaaccat 1200 acggatcatt tctcagtagc catattcacc atccaccgga tcctcccocc tgactcagga 1260 gtttgggtct gcagtgtgaa cacagtggct gggatggtgg aaaagccctt caaoatttct 1320 gttaaagttc ttccaaagcc cctgaatgcc ccaaacgtga ttgacactgg acataacttt 1380 gctgtcatca acatcagctc tgagcqttac tttggggatg gaccaatcaa atccaagaag 1440 qttetataea aacccgttaa tcactatgag gcttggcaac atattcaagt gagaaatgag 1500 attgttaCac tqaaqtattt ggaaqqtqgg aqagaatatg aaetetgtgt gcaaCtggtc 1560 qgtegtggag agggtgggga agggeatect ggaectgtga gacgcttcac aacagcttct 1620 atcggactcc ctcctccaag aggtetaaat ctcctgccta aaagtcagac cactctaaat 1680 ttgacctggc aaccaatatt tccaagctcg gaagatgact tttatgttga agtggagaga 1740 aggtctgtgc aaaaaagtga tcagcagaat attaaagttc caggcaactt gacttcggtg 1800 ctacttaaca acttacatcc cagggagcag tacgtggtcc gagctagagt caacaccaag 1860 gcccaggggg aatggagtga agatctcact gcttggaccc ttagtgacat tcttcctcct 1920 caaccagaaa acatcaagat ttccaacatt acacactcct cggctgtgat ttcttggaca 1980 atattggatg gctattctat ttcttctatt actatccgtt acaaggttca aggcaagaat 2040 gaagaccagc acgttgatgt gaagataaag aatgccacca tcattcagta tcagctcaag 2100 ggcctagagc ctgaaacagc ataccaggtg gacatttttg cagagaacaa catagggtca 2160 agcaacccag Ccttttctca tgaactggtg accctcccag aatctcaagc accagcggac 2220 ctcggagggg ggaagatgct gcttatagcc atccttggct ctgctggaat gacctgcctg 2280 actgtgctgt tggcctttct gatcatattg caattgaaga gggcaaatgt gcaaaggaga 2340 atggcccaag ccttccaaaa cgtgagggaa gaaccagctg tgcagttcaa ctcagggact 2400 ctggccctaa acaggaaggt caaaaacaac ecagatccta caatttatcc agtgcttgac 2460 tggaatgaca tcaaatttca agatgtgatt ggggagggca attttggcca agttcttaag 2520 gcgcgcatca agaaggatgg gttacggatg gatgctgcoa tcaaaagaat gaaagaatat 2580 gcctccaaag atgatcacag ggactttgca ggagaactgg aagttctttg taaacttgga 2640 caccatccaa aCatCatcaa tctcttagga gcatgtgaac atcgaggcta Cttgtacctg 2700 gccattgagt acgcgcccCa tggaaacctt ctggacttcc ttcgcaagag ccgtgtgctg 2760 gagacggacc cagcatttgc cattgccaat agcaccgcgt ccacactgtc ctcccagcag 2820 ctccttcact tcgctgccga cgtggcccgg ggcatggact acttgagcca aaaacagttt 2880 atccacaggg atctggctgc cagaaacatt ttagttggtg aaaactatgt ggcaaaaata 2940 gcagattttg gattgtcccg aggtcaagag gtgtacgtga aaaagacaat gggaaggctc 3000 ccagtgcgct ggatggccat cgagtcactg aattacagtg tgtacacaac caacagtgat 3060 gtatggtcct atggtgtgtt actatgggag attgttagct taggaggcac accCtactgc 3120 gggatgactt gtgcagaact ctacgagaag ctgccccagg gctacagact ggagaagccc 3180 ctgaactgtg atgatgaggt gtatgatcta atgagacaat gctggcggga gaagccttat 3240 gagaggccat catttgccca gatattggtg tccttaaaca gaatgttaga ggagcgaaag 3300 acctacgtga ataccacgct ttatgagaag tttacttatg caggaattga ctgttctgct 3360 gaagaagcgg cctag 3375 <210> 183 <211 >4654

<212> DNA <213> Mouse <400> 183 gagcaggagc cggagcagga gcagaagata agccttggat gaagggcaag atggataggg 60 ctcgctctgc cccaagccct gctgatacca agtgccttta agatacagcc tttcccatcc 120 taatctgcaa aggaaacagg aaaaaggaac ttaaccctcc ctgtgctcag acagaaatga 180 gactgttacc gcctgcttct gtggtgtttc tccttgccgc caacttgtaa acaagagcga 240 gtggaccatg cgagcgggaa gtcgcaaagt tgtgagttgt tgaaagcttc CcagggaCtc 300 atgctcatct gtggacgctg gatggggaga tctggggaag tatggactct ttagccggct 360 tagttctctg tggagtcagc ttgctccttt atggagtagt agaaggcgcc atggacctga 420 tcttgatcaa ttccctacct cttgtgtctg atgccgaaac atccctcacc tgcattgcct 480 ctgggtggca cccccatgag cccatcacca taggaaggga ctttgaagcc ttaatgaacc 540 agcaccaaga tccactggag gttactcaag atgtgaecag agaatgggcg aaaaaagttg 600 tttggaagag agaaaaggcc agtaagatta atggtgctta tttctgtgaa ggtcgagttc 660 gaggacaggc tataaggata cggaccatga agatgcgtca acaagcatcq ttcctacctg 720 ctactttaac tatgaccgtg gacaggggag ataatgtgaa catatctttc aaaaaggtgt 780 taattaaaga agaagatgca gtgatttaca aaaatggctc cttcatccac tcagtgcccc 840 ggcatgaagt acctgatatt ttagaagttc acttgccgca tgctcagcoc caggatgctg 900 gtgtgtactc ggccaggtac ataggaggaa acctgttcac ctcagccttc accaggctga 960 ttgttcggag atgtgaagct cagaagtggg ggcccgactg tagccgtcct tgtactactt 1020 gcaagaacaa tggagtctgc catgaagata ccggggaatg catttgecct cctgggttta 1080 tggggagaac atgtgagaaa gcttgtgagc cgcacacatt tggcaggaoc tgtaaagaaa 1140 ggtgtagtgg accagaagga tgcaagtctt atgtgttctg tctcccagac ccttacgggt 1200 gttcctgtgc cacaggctgg agggggttgc agtgcaatga agcatgccca tctggttact 1260 acggacaaga ctgtaagctc aggtgccact gtaocaatga agagatatgt gatcggttcc 1320 aaggatgcct ctgctctcaa ggatggcaag ggctgaagtg tgagaaagaa ggaaggccaa 1380 ggatgactcc acagatagag gatttgccag atcacattga agtaaacagt ggaaaattta 1440 accccatctg caaagcctct gggtggccac tacctactag tgaagaaatg accctagtga 1500 agccagatgg gacagtgctc caaccaaatg acttcaacta tacagatcgt ttctcagtgg 1560 ccatattcac tgtcaaccga gtcttacctc ctgactcagg agtctgggtc tgcagtgtga 1620 acacagtggc tgggatggtg gaaaagcctt tcaacatttc cgtcaaagtt cttccagagc 1680 ccctgcacgc cccaaatgtg attgacactg gacataactt tgctatcatc aatatcagct 1740 ctgagcctta ctttggggat ggacccatca aatccaagaa gcttttctat aaacctgtca 1800 atcaggcctg gaaatacatt gaagtgacga atgagatttt cactotcaac tacttggagc 1860 cgcggactga ctacgagctg tgtgtgcagc tggcccgtcc tggagagggt ggagaagggc 1920 atcctgggcc tgtgagacga tttacaacag cgtctatcgg actccctcct ccaagaggtc 1980 tcagtctcct gccaaaaagc cagacagctc taaatttgac ttggcaaccg atatttacaa 2040 actcagaaga tgaattttat gtggaagtcg agaggcgatc cctgcaaaca acaagtgatc 2100 agcagaacat caaagtgcct gggaacctga cctcggtgct actgagcaac ttagtcccca 2160 gggagcagta cacagtccga gctagagtca acaccaaggc gcagggggag tggagtgaag 2220 aactcagggc ctggaccctt agtgacattc tccctcctca accagaaaac atcaagatct 2280 ccaacatCac tgactccaca gctatggttt cttggacaat agtggatggc tattcgattt 2340 cttccatcat catccggtat aaggttcagg gcaaaaatga agaccagcae attgatgtga 2400 agatcaagaa tgctaqcgtt actcagtacc agctcaaggg cctagagcca gagactacat 2460 accatgtgga tatttttgct gagaacaaca taggatcaag caacccagcc ttttctcatg 2520 aaotgaggac gcttccacat tccccagcct ctgcagacct cggaggggga aagatgctac 2580 tcatagccat ccttgggtcg gctggaatga cttgcatcac cgtgctgttg gcgtttctga 2640 ttatgttgca actgaagaga gcaaatgtcc aaaggagaat ggctcaggca ttccagaacg 2700 tgagagaaga accagctgtg cagtttaact caggaactct ggcccttaac aggaaggcca 2760 aaaacaatcc ggatcccaca atttatcctg tgcttgactg gaatgacatc aagtttcaag 2820 acgtgatcgg agagggcaac tttggccagg ttctgaaggc acgcatcaag aaggatgggt 2880 tacggatgga tgcegccatc aagaggatga aagagtatgc ctccaaagat gatcacaggg 2940 acttcgcagg agaactggag gttctttgta aacttggaca ccatccaaac ateattaatc 3000 tcttgggagc atgtgaacac cgaggctatt tgtacctagc tattgagtat gccccgcatg 3060 gaaacctcct ggacttcctg cgtaagagca gagtgctaga gacagaccct gcttttgcca 3120 tcgccaacag tacagcttcc acactgtcct cccaacagct tcttcatttt gctgcagatg 3180 tggcccgggg gatggactac ttgagccaga aacagtttat Ccacagggac ctggctgcca 3240 gaaacatttt agttggtgaa aactacatag ccaaaatagc agattttgga ttgtcacgag 3300 gtcaagaagt gtatgtgaaa aagacaatgg gaaggctccc agtgcgttgg atggcaatcg 3360 aatcactgaa ctatagtgtc tatacaacca acagtgatgt ctggtcctat ggtgtattgc 3420 tctgggagat tgttagctta ggaggcaccc cctactgcgg catgacgtgc gcggagctct 3480 atgagaagct accccagggc tacaggctgg agaagcccct gaactgtgat gatgaggtgt 3540 atgatctaat gagacagtgc tggagggaga agccttatga gagaccatca tttgcccaga 3600 tattggtgtc cttaaacagg atgctggaag aacggaagac atacgtgaac accacactgt 3660 atgagaagtt tacctatgca ggaattgact gctctgcgga agaagcagcc tagagcagaa 3720 ctcttcatgt acaacggcca tttctcctca ctggcgcgag agcgccttga cacctgtacc 3780 aagcaagcca cccactgcca agagatgtga tatataagtg tatatattgt gctgtgtttg 3840 ggaccctcct catacagctc gtgcggatct gcagtgtgtt ctgactctaa tgtgactgta 3900 tatactgctc ggagtaagaa tgtgctaaga tcagaatgcc tgttcgtggt ttcatataat 3960 atatttttct aaaagcatag attgcacagg aaggtatgag tacaaatact gtaatgcata 4020 acttgttatt gtcctagatg tgtttgatat ttttccttta caactgaatg ctataaaagt 4080 gttttgctgt gtacacataa gatactgttc gttaaaataa gcattccctt gacagcacag 4140 gaagaaaago gagggaaatg tatggattat attaaatgtg ggttactaca caagaggccg 4200 aacattccaa gtagcagaag agagggtctc tcaactctgc tcotcacotg cagaagccag 4260 tttgtttggc Catgtgacaa ttgtcctgtg tttttatagc acccaaatca ttctaaaata 4320 tgaacatcta aaaactttgc taggagacta agaacctttg gagagataga tataagtacg 4380 gtcaaaaaac aaaactgtgg gacttacatt tattttctat agtaatctgt tgtacatttt 4440 aagaagtaaa actaggaatt taggagtgat gtgtgacatt tctgacatgg agttaccatc 4500 cccacatgta tcacatactg tcatattccc acatgtatca cacatgtatt gtaaaatttt 4560 gtagttttga tcacttgtga atttactgtt gatgtggtag ccacctgctg caatggttcc 4620 tcttgtaggt gaataaatgt cttgtctacc caca 4654 <210> 184 <211> 3378

<212> DNA <213> Artificial <220> <223> Mutated Cyno sequence <400> 184 atggacagcc tggccagcct ggtgctgtgc ggagtgtccc tgctgctgag cggcacagtg 60 gaaggcgcca tggacctgat cctgatcaac tccctgcccc tggtgtccga cgccgagaca 120 agcctgacct gtatcgctag cggctggcac ccccacgagc ctatcaccat cggccgggac 180 ttcgaggccc tgatgaacca gcaccaggac cccctggaag tgacccagga cgtgacccgc 240 gagtgggeca agaaagtcgt gtggaagaga gagaaggcca gcaagatcaa cggcgcctac 300 ttctgcgagg gcagagtgcg gggcgaggcc atcagaatcc ggacaatgaa gatgcggcag 360 caggccagct tcctgcqcgc caccctgacc atgaccgtgg acaagggcga caacgtgaac 420 atcagcttca agaaggtgct gatcaaagaa gaggacgccg tcatctacaa gaacggcagc 480 ttcatccaca gcgtgccccg gcacgaggtg cccgacatcc tggaagtgca tctgccccac 540 gcccagcctc aggatgccgg cgtgtacagc gcccggtaca tcggcggcaa cctgttcacc 600 agcgccttca Cccggctgat cgtgcggaga tgcgaggccc agaaatgggg ccctgagtgt 660 aaccggctgt gcaccgtgtg cgtgaacaac ggcgtgtgcc acgaggacac cggcgagtgc 720 atctgtcctc ccggcttcat gggccggacc tgcgagaagg cctgcgagag acacaccttc 780 ggcagaacct gcaaagagcg gtgtagcggc caggacggct gcaagagCta cgtgttctgc 840 ctgcccgacc cctacggctg cagctgtgcc acaggctgga agggcctgca gtgcaacgag 900 gcctgccacc acggcttcta cggccccgac tgcaagctgc ggtgcagctg cagcaacggc 960 gagacatgcg accggttcca gggatgcctg tgcagccctg gcagacaggg actccagtgc 1020 gagcgcgagg gcatcccqag aatgaccccc aagatcgtgg acctgcccga tcacatcgaa 1080 gtgaacagcg gcaagttcaa ccccatctgc aaggcctccg gctggcccct gcccaccaac 1140 gaagagatga ccctggteaa gcccgacggc accgtgctgc accccaagga cttcaaccac 1200 accgaccact tcagcgtggc catcttcacc atccaccgga tcctgccccc cgactctggc 1260 gtgtgggtct gcagcgccaa taccgtggcc ggcatggtgg aaaagccctt caacatcagc 1320 gtgaaggtgc tgcccaagcq cctgaacgaa cccaaagtga tcgacaacgg acacaacttc 1380 gccgtgatca acatctccag cgagccctac ttcggcgacg gccccatcaa gagcaagaag 1440 ctgctgtaca agcccgtgaa ccactacgag gcctggcggc acatccaggt cacaaacgag 1500 atcgtgaccc tgaaccacct ggaaccccgg accgagtacg agctgtgcgt gcagctcgtg 1560 cggaggggcg aaggcggcga aggacatcct ggccccgtgc gcagattcac caccgcctct 1620 atcggcctgc ccccacccag aggcctgaac ctgctgccta agagccagac caccctgaac 1680 ctgacctggc agcccatctt ccccagcagc gaggacgact tctacgtgga agtggaacgg 1740 cggagcgtgc agaagtccga ccagcagaac atcaaggtgc ccggcaacct gaccagcgtg 1800 ctgctgaaca acctgcaccc ccgcgagcag tacgtcgtgc gggccagagt gaacaccaag 1860 gcccagggcg agtggtccga ggatctgacc gcctggaccc tgagcgacat cctgcctccc 1920 cagcccgaga atatcaaaat cagcaacatc acccacagea gcgcegtgat ctcttggacc 1980 atcctggacg gctacagcat cagctccatc accatccggt acaaggtgca gggcaagaac 2040 gaggaccagc acatcgacgt gaagatcaag aacgccacca tcacccagta ccagctgaag 2100 ggcctggaac ccgagacagc ctaccaggtg gacatcttcg ccgagaacaa tatcggcagc 2160 agcaaccccg ccttcagcca cgagctggtc accctgcccg agtctgaggc ccctgctgat 2220 ctgggcggag gcaagatgct gctgatcgcc atcctgggca gcgccggcat gacctgtctg 2280 acagtgctgc tggccttcct gatcatcctg cagctgaagc gggccaacgt gcagcggaga 2340 atggcccagg ccttccagaa cgtgcgcgag gaacccgccg tgcagttcaa cagcggcacc 2400 ctggccctga accggaaagt gaagaacaac cccgacccca ccatctaccc cgtgctggac 2460 tggaacgaca tcaagttcca ggatgtgatc ggcgagggca acttcggcca ggtgctgaag 2520 gcccggatca agaaagacgg cctgcggatg gacgccgcca tcaagcggat gaaggaatac 2580 gccagcaagg acgacqacag agacttcgcc ggggagctgg aagtgctgtg caagctgggc 2640 caccacccca acatcatcaa cctgctgggc gcctgcgagc accggggcta tctgtacctg 2700 gccatcgagt acgcccccca cggcaatctg ctggatttcc tgcggaagtc ccgggtgctg 2760 gaaaccgacc ctgccttcgc cattgccaac agcaccgcca gcaccctgag cagccagcag 2820 ctcctgcact tcgccgccga tgtggccaga ggcatggact acctgagcca gaagcagttc 2880 atccaccggg acctggccgc cagaaacatc ctcgtgggcg agaactacgt ggccaagatc 2940 gccgacttcg gcctgagcag aggccaggaa gtgtacgtca agaaaaccat gggccggctg 3000 cccgtgcggt ggatggccat tgagagcctg aactacagcg tgtacaccac caacagcgac 3060 gtgtggtcct acggggtgct gctgtgggag atcgtgtetc tgggcggcac cccctactgc 3120 ggcatgacat gcgccgagct gtacgagaag ctgccccagg gctaccggct ggaaaagcct 3180

Ctgaactgcg acgacgaggt gttcgacctg atgcggcagt gctggcgcga gaagccctac 3240 gagcggccta gcttcgccca gatcctggtg tccctgaaca gaatgctgga agaacgcaag 3300 acctacgtga acaccaccct gtatgagaag ttcacctacg ccggcatcga ctgcagcgcc 3360 gaagaggccg cttgatga 3378 <210> 185 <211> 1491

<212> DNA <213> Mouse <400> 185 atgtggcaga tcattttcct aacttttggc tgggatcttg tcttggcctc agcqtacagt 60 aactttagga agagcgtgga cagcacaggc agaaggcagt accaggtcca gaacggaccc 120 tgcagctaca cgttcqtgct gccggagacc gacagctgcc gatcttcctc cagcccctac 180 atgtccaatg ccgtgcagag ggatgcaccc ctcgactacg acgactcagt gcaaaggctg 240 caggtgctgg agaacattct agagaacaac acacagtggc tgatgaagct ggagaattac 300 attcaggaca acatgaagaa ggagatggtg gagatccaac agaatgtggt gcagaaccag 360 acagctgtga tgatagagat tggaaccagc ttgctgaacc agacagcagc acaaactcgg 420 aaactgactg atgtggaagc ccaagtacta aaccagacga caagactcga gctgcagctt 480 ctccaacatt ctatttctac caacaaattg gaaaagcaga ttttggatca gaccagtgaa 540 ataaacaagc tacaaaataa gaacagcttc ctagaacaga aagttctgga catggagggc 600 aagcacagcg agcagctaca gtccatgaag gagcagaagg acgagctcca ggtgctggtg 660 tccaagcaga gctctgtcat tgacgagctg gagaagaagc tggtgacagc cacggtcaac 720 aactcgctcc ttcagaagca gcagcatgac ctaatggaga ccgtcaacag cttgctgacc 780 atgatgtcat cacccaactc caagagctcg gttgctatcc gtaaagaaga gcaaaccacc 840 ttcagagact gtgcggaaat cttcaagtca ggactcacca ccagtggcat ctacacactg 900 accttcccca actccacaga ggagatcaag gcctactgtg acatggacgt gggtggagga 960 gggtggacag tcatccaaca ccgagaagat ggcagtgtgg acttccagag gacgtggaaa 1020 gaatacaaag agggcttcgg gagccctctg ggagagtact ggctgggcaa tgagtttgtc 1080 tcccagctga ccggtcagca ccgctacgtg cttaagatcc agctgaagga ctgggaaggc 1140 aacgaggcgc attcgctgta tgatcacttc tacctcgctg gtgaagagtc caactacagg 1200 attcacctta caggactcac ggggaccgcg ggcaaaataa gtagcatcag ccaaccagga 1260 agtgatttta gcacaaagga ttcggacaat gacaaatgca tctgcaagtg ttaccagatg 1320 ctctcaggag gctggtggtt tgacgcatgt ggtccttcca acttgaatgg acagtactac 1380 ccacaaaaac agaatacaaa taagtttaac ggtatcaagt ggtactactg gaaggggtcc 1440 ggctactcgc tcaaggccac aaccatgatg atccggccag cagatttcta a 1491 <210> 186 <211> 1488

<212> DNA <213> Cyno <400> 186 atgtggcaga ttgttttctt taccctgagc tgtgatcttg tcttggccgc agcctataac 60 aactttcgga agagcatgga cagcatagga aagaagcagt atcaggtcca gcatgggtcc 120 tgcagctaca ctttcctcct gccagagatg gacaactgcc gctcttcctc CagcCcctac 180 gtgtccaatg ctgtgcagag ggacgcgccg ctagaatatg atgactcggt gcagaggctg 240 caagtgctgg agaacatcat ggaaaacaac actcagtggc taatgaagct tgagaattat 300 atccaggaca acatgaaaaa agaaatggta gagatacagc agaacgcagt acagaaccag 360 acggctgtga tgatagaagt agtgacgaac ctgttgaacc aaacggcgga gcaaacgcgg 420 aagttaactg atgtggaagc ccaagtatta aaccagacca caagacttga acttcagctc 480 ttggaacact Ccctctcgac aaacaaattg gaaaaacaga ttttggacca gaccagtgaa 540 ataaacaaat tgcaagataa gaacagtttc ctagaaaaga aggtgctagc tatggaagac 600 aagcacatca tccaactaca gtcaatcaaa gaagagaaag atcagctgca ggtgttagta 660 tccaagcaaa attccatcat tgaagaacta gaaaaaaaaa tagtgactgc cacggtgaat 720 aattcagttc ttcagaagca gcaacatgat ctcatggaga cagttaataa cttactgact 780 atgatgtcca catcaaactc taaggacccc actgttgcta aggaagaaca aatcagcttc 840 agagactgtg ctgaagtgtt caaatcagga cacaccacga atggcgtcta cacgttaaca 900 ctccccaact ctacagaaga ggtcaaggcc tactgtgaca tggaagctgg tggaggcggg 960 tggacaatta ttcagcgacg tgaggacggc agcgttgact ttcagaggac ttggaaagaa 1020 tataaagtgg gatttggtaa cccttcagga gaatattggc tgggaaatga gtttgtttca 1080 caactgacta atcagcaacg ctatgtgctt aaaatacacc ttaaagactg ggaagggaac 1140 gaggcttact cattgtatga acatttctat ctctcaagtg aagaactCaa ttataggatt 1200 caccttaaag gacttacagg gacagccggc aaaataagca gcatcagtca accaggaaat 1260 gattttagca caaaggatgc agacaacgac aaatgtattt gcaaatgttc acaaatgcta 1320 acaggaggct ggtggtttga tgcatgtggt ccttccaact tgaacggaat gtactatcca 1380 caaaggcaga acacaaataa gttcaacggc attaagtggt actactggaa aggctcaggc 1440 tactcgctca agggcacaac catgatgatc cgaccggcag atttctaa 1488 <210> 187 <211> 495

<212> PRT <213> Cyno <400> 187

Met Trp Gin lie Val Phe Phe Thr Leu Ser Cys Asp Leu Val Leu Ala 15 10 15

Ala Ala Tyr Asn Asn Phe Arg Lys Ser Met Asp Ser He Gly Lys Lys 20 25 30

Gin Tyr Gin Val Gin His Gly Ser Cys Ser Tyr Thr Phe Leu Leu Pro 35 40 45

Glu Met Asp Asn Cys Arg Ser Ser Ser Ser Pro Tyr Val Ser Asn Ala 50 55 60

Val Gin Arg Asp Ala Pro Leu Glu Tyr Asp Asp Ser Val Gin Arg Leu 65 70 75 80

Gin Val Leu Glu Asn lie Met Glu Asn Asn Thr Gin Trp LeU Met Lys 85 90 95

Leu Glu Asn Tyr He Gin Asp Asn Met Lys Lys Glu Met Val Glu He 100 105 110

Gin Gin Asn Ala Val Gin Asn Gin Thr Ala Val Met He Glu Val Val 115 120 125

Thr Asn Leu Leu Asn Gin Thr Ala Glu Gin Thr Arg Lys Leu Thr Asp 130 135 140

Val Glu Ala Gin Val Leu Asn Gin Thr Thr Arg Leu Glu Leu Gin Leu 145 150 155 160

Leu Glu His Ser Leu Ser Thr Asn Lys Leu Glu Lys Gin lie Leu Asp 165 170 175

Gin Thr Ser Glu lie Asn Lys Leu Gin Asp Lys Asn Ser Phe Leu Glu 180 185 190

Lys Lys Val Leu Ala Met Glu Asp Lys His lie lie Gin Leu Gin Ser 195 200 205

He Lys Glu Glu Lys Asp Gin Leu Gin Val Leu Val Ser Lys Gin Asn 210 215 220

Ser He lie Glu Glu Leu Glu Lys Lys He Val Thr Ala Thr Val Asn 225 230 235 240

Asn Ser Val Leu Gin Lys Gin Gin His Asp Leu Met Glu Thr Val Asn 245 250 255

Asn Leu Leu Thr Met Met Ser Thr Ser Asn Ser Lys Asp Pro Thr Val 260 265 270

Ala Lys Glu Glu Gin He Ser Phe Arg Asp Cys Ala Glu Val Phe Lys 275 280 285

Ser Gly His Thr Thr Asn Gly Val Tyr Thr Leu Thr Leu Pro Asn Ser 290 295 300

Thr Glu Glu Val Lys Ala Tyr Gys Asp Met Glu Ala Gly Gly Gly Gly 305 310 315 320

Trp Thr He He Gin Arg Arg Glu Asp Gly Ser Val Asp Phe Gin Arg 325 330 335

Thr Trp Lys Glu Tyr Lys Val Gly Phe Gly Asn Pro Ser Gly GlU Tyr 340 345 350

Trp Leu Gly Asn Glu Phe Val Ser Gin Leu Thr Asn Gin Gin Arg Tyr 355 360 365

Val Leu Lys He His Leu Lys Asp Trp Glu Gly Asn Glu Ala Tyr Ser 370 375 380

Leu Tyr Glu His Phe Tyr Leu Ser Ser Glu Glu Leu Asn Tyr Arg He 385 390 395 400

His Leu Lys Gly Leu Thr Gly Thr Ala Gly Lys He Ser Ser He Ser 405 410 415

Gin Pro Gly Asn Asp Phe Ser Thr Lys Asp Ala Asp Asn Asp Lys Cys 420 425 430

He Cys Lys Cys Ser Gin Met Leu Thr Gly Gly Trp Trp Phe Asp Ala 435 440 445

Cys Gly Pro Ser Asn Leu Asn Gly Met Tyr Tyr Pro Gin Arg Gin Asn 450 455 460

Thr Asn Lys Phe Asn Gly lie Lys Trp Tyr Tyr Trp Lys Gly Ser Gly 465 470 475 480

Tyr Ser Leu Lys Gly Thr Thr Met Met lie Arg Pro Ala Asp Phe 485 490 495 <210> 188 <211> 496

<212> PRT <213> Mouse <400> 188

Met Trp Gin He He Phe Leu Thr Phe Gly Trp Asp Leu Val Leu Ala 15 10 15

Ser Ala Tyr Ser Asn Phe Arg Lys Ser Val Asp Ser Thr Gly Arg Arg 20 25 30

Gin Tyr Gin Val Gin Asn Gly Pro Cys Ser Tyr Thr Phe Leu Leu Pro 35 40 45

Glu Thr Asp Ser Cys Arg Ser Ser Ser Ser Pro Tyr Met Ser Asn Ala 50 55 60

Val Gin Arg Asp Ala Pro Leu Asp Tyr Asp Asp Ser Val Gin Arg Leu 65 70 75 80

Gin Val Leu Glu Asn He Leu Glu Asn Asn Thr Gin Trp Leu Met Lys 85 90 95

Leu Glu Asn Tyr He Gin Asp Asn Met Lys Lys Glu Met Val Glu He 100 105 110

Gin Gin Asn Val Val Gin Asn Gin Thr Ala Val Met He Glu He Gly 115 120 125

Thr Ser Leu Leu Asn Gin Thr Ala Ala Gin Thr Arg Lys Leu Thr Asp 130 135 140

Val Glu Ala Gin Val Leu Asn Gin Thr Thr Arg Leu Glu Leu Gin Leu 145 150 155 160

Leu Gin His Ser He Ser Thr Asn Lys Leu Glu Lys Gin He Leu Asp 165 170 175

Gin Thr Ser Glu He Asn Lys Leu Gin Asn Lys Asn Ser Phe Leu Glu 180 185 190

Gin Lys Val Leu Asp Met Glu Gly Lys His Ser Glu Gin Leu Gin Ser 195 200 205

Met Lys Glu Gin Lys Asp Glu Leu Gin Val Leu Val Ser Lys Gin Ser ζιυ 215 ζζυ

Ser val lie Asp Glu Leu Glu Lys Lys Leu Val Thr Ala Thr Val Asn 225 230 235 240

Asn Ser Leu Leu Gin Lys Gin Gin His Asp Leu Met Glu Thr Val Asn 245 250 255

Ser Leu Leu Thr Met Met Ser Ser Pro Asn Ser Lys Ser Ser Val Ala 260 265 270 lie Arg Lys Glu Glu Gin Thr Thr Phe Arg Asp Cys Ala Glu lie phe 275 280 285

Lys Ser Gly Leu Thr Thr Ser Gly Tie Tyr Thr Leu Thr Phe Pro Asn 290 295 300

Ser Thr Glu Glu He Lys Ala Tyr Cys Asp Met Asp Val Gly Gly Gly 305 310 315 320

Gly Trp Thr Val He Gin His Arg Glu Asp Gly Ser Val Asp Phe Gin 325 330 335

Arg Thr Trp Lys Glu Tyr Lys G1U Gly Phe Gly Ser Pro Leu Gly Glu 340 345 350

Tyr Trp Leu Gly Asn Glu Phe Val Ser Glh Leu Thr Gly Gin His Arg 355 360 365

Tyr Val Leu Lys He Gin Leu Lys Asp Trp Glu Gly Asn Glu Ala His 370 375 380

Ser Leu Tyr Asp His Phe Tyr Leu Ala Gly Glu Glu Ser Asn Tyr Arg 385 390 395 400 lie His Leu Thr Gly Leu Thr Gly Thr Ala Gly Lys He Ser Ser He 405 410 415

Ser Gin Pro Gly Ser Asp Phe Ser Thr Lys Asp Ser Asp Asn Asp Lys 420 425 430

Cys lie Cys Lys Cys Ser Gin Met Leu Ser Gly Gly Trp Trp Phe Asp 435 440 445

Ala Cys Gly Pro Ser Asn Leu Asn Gly Gin Tyr Tyr Pro Gin Lys Gin 450 455 460

Asn Thr Asn Lys Phe Asn Gly He Lys Trp Tyr Tyr Trp Lys Gly Ser 465 470 475 480

Gly Tyr Ser Leu Lys Ala Thr Thr Met Met He Arg Pro Ala Asp Phe 485 490 495 <210> 189 <211> 253

<212> PRT <213> Mouse <400> 189

Met Glu Thr Asp Thr Leu Leu Leu Trp Val Leu Leu Leu Trp Val Pro 15 10 15

Gly Ser Thr Gly Asp Asp Tyr Lys Asp Asp Asp Asp Lys Gly lie Leu 20 25 30

Glu Glu Gin Thr Thr Phe Arg Asp Cys Ala Glu He Phe Lys Ser Gly 35 40 45

Leu Thr Thr Ser Gly He Tyr Thr Leu Thr Phe Pro Asn Ser Thr Glu 50 55 60

Glu He Lys Ala Tyr Cys Asp Met Asp Val Gly Gly Gly Gly Trp Thr 65 70 75 80

Val He Gin His Arg Glu Asp Gly Ser Val Asp Phe Gin Arg Thr Trp 85 90 95

Lys Glu Tyr Lys Glu Gly Phe Gly Ser Pro Leu Gly Glu Tyr Trp Leu 100 105 110

Gly Asn Glu Phe Val Ser Gin Leu Thr Gly Gin His Arg Tyr Val Leu 115 120 125

Lys He Gin Leu Lys Asp Trp Glu Gly Asn Glu Ala His Ser Leu Tyr 130 135 140

Asp His Phe Tyr Leu Ala Gly Glu Glu Ser Asn Tyr Arg He His Leu 145 150 155 160

Thr Gly Leu Thr Gly Thr Ala Gly Lys He Ser Ser He Ser Gin Pro 165 170 175

Gly Ser Asp Phe Ser Thr Lys Asp Ser Asp Asn Asp Lys Cys He Cys 180 185 190

Lys Cys Ser Gin Met Leu Ser Gly Gly Trp Trp Phe Asp Ala Cys Gly 195 200 205

Pro Ser Asn Leu Asn Gly Gin Tyr Tyr Pro Gin Lys Gin Asn Thr Asn 210 215 220

Lys Phe Asn Gly He Lys Trp Tyr Tyr Trp Lys Gly Ser Gly Tyr Ser 225 230 235 240

Leu Lys Ala Thr Thr Met Met He Arg Pro Ala Asp Phe 245 250 <210> 190 <211> 253

<212> PRT <213> Cyno <400> 190

Met Glu Thr Asp Thr Leu Leu Leu Trp Val Leu Leu Leu Trp Val Pro 15 10 15

Gly Ser Thr Gly Asp Asp Tyr Lys Asp Asp Asp Asp Lys Gly lie Leu 20 25 30

Glu Glu Gin He Ser Phe Arg Asp Cys Ala Glu Val Phe Lys Ser Gly 35 40 45

His Thr Thr Asn Gly Val Tyr Thr Leu Thr Leu Pro Asn Ser Thr Glu 50 55 60

Glu Val Lys Ala Tyr Cys Asp Met Glu Ala Gly Gly Gly Gly Trp Thr 65 70 75 80

He He Gin Arg Arg Glu Asp Gly Ser Val Asp Phe Gin Arg Thr Trp 85 90 95

Lys Glu Tyr Lys Val Gly Phe Gly Asn Pro Ser Gly Glu Tyr Trp Leu 100 105 110

Gly Asn Glu Phe Val Ser Gin Leu Thr Asn Gin Gin Arg Tyr Val Leu 115 120 125

Lys He His Leu Lys Asp Trp Glu Gly Asn Glu Ala Tyr Ser Leu Tyr 130 135 140

Glu His Phe Tyr Leu Ser Ser Glu Glu Leu Asn Tyr Arg He His Leu 145 150 155 160

Lys Gly Leu Thr Gly Thr Ala Gly Lys He Ser Ser He Ser Gin Pro 165 170 175

Gly Asn Asp Phe Ser Thr Lys Asp Ala Asp Asn Asp Lys Cys lie Cys 180 185 190

Lys Cys Ser Gin Met Leu Thr Gly Gly Trp Trp Phe Asp Ala Cys Gly 195 200 205

Pro Ser Asn Leu Asn Gly Met Tyr Tyr Pro Gin Arg Gin Asn Thr Asn 210 215 220

Lys Phe Asn Gly He Lys Trp Tyr Tyr Trp Lys Gly Ser Gly Tyr Ser 225 230 235 240

Leu Lys Gly Thr Thr Met Met He Arg Pro Ala Asp Phe 245 250

REFERENCES CITED IN THE DESCRIPTION

This list of references cited by the applicant is for the reader's convenience only. It does not form part of the European patent document. Even though great care has been taken in compiling the references, errors or omissions cannot be excluded and the EPO disclaims all liability in this regard.

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Claims (15)

An Ang2 binding molecule comprising a single variable immunoglobulin domain, wherein the single variable immunoglobulin domain comprises three complementarity determining regions CDR1, CDR2 and CDR3, wherein CDR1 has an amino acid sequence shown in SEQ ID NO: 168, CDR2 has an amino acid sequence shown in SEQ ID NO. : 171, and CDR3 has an amino acid sequence shown in SEQ ID NO: 175. The Ang2 binding molecule of claim 1, wherein the single variable immunoglobulin domain is a VHH. The Ang2 binding molecule of claim 2, wherein said VHH consists of a single variable immunoglobulin domain having a sequence consisting of SEQ ID NO: 166. An Ang2 binding molecule according to any one of claims 1 to 3, wherein said VHH or said single variable immunoglobulin domain has a modification or replacement at its N-terminal end, wherein the modification is a deletion of the first amino acid and the replacement is a replacement of the first amino acid with a second amino acid. A nucleic acid molecule encoding the Ang2 binding molecule according to any one of claims 1 to 4. An expression vector comprising the nucleic acid molecule of claim 5. A host cell carrying one or more expression vectors according to claim 6. A method of producing the Ang2 binding molecule according to any one of claims 1 to 4, comprising the steps of: transfecting a host cell with one or more of the expression vectors of claim 6, culturing the host cell, and isolating and purifying the Ang2-binding molecule. A pharmaceutical composition comprising as active ingredient one or more of said Ang2-binding molecules according to any one of claims 1 to 4 and at least one physiologically acceptable carrier. The pharmaceutical composition of claim 9, further comprising one or more additional therapeutic agents. A pharmaceutical composition according to any one of claims 9 or 10 for use in the treatment of a disease associated with Ang2-mediated effects on angiogenesis. The pharmaceutical composition of claim 11 for use in the treatment of cancer and cancer diseases. The pharmaceutical composition of claim 11 for use in the treatment of eye diseases. The pharmaceutical composition of claim 13, wherein the eye disease is age-related macular degeneration or diabetic retinopathy. The pharmaceutical composition of claim 11 for use in the treatment of chronic kidney disease.